==== Front
Curr Res Neurobiol
Curr Res Neurobiol
Current Research in Neurobiology
2665-945X
Elsevier

S2665-945X(23)00014-1
10.1016/j.crneur.2023.100086
100086
Articles from the special issue: Illuminating the Monkey Brain: Organization, Networks and Circuits, edited by Yogita Chudasama and Xiaoqin Wang
Advances in AAV technology for delivering genetically encoded cargo to the nonhuman primate nervous system
Campos Lillian J. ac1
Arokiaraj Cynthia M. bc1
Chuapoco Miguel R. bc
Chen Xinhong b
Goeden Nick bd
Gradinaru Viviana viviana@caltech.edu
bc∗
Fox Andrew S. dfox@ucdavis.edu
ac∗∗
a Department of Psychology and the California National Primate Research Center, University of California, Davis, CA, 05616, USA
b Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, 91125, USA
c Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815, USA
d Capsida Biotherapeutics, Thousand Oaks, CA, 91320, USA
∗ Corresponding author. Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, 91125, USA. viviana@caltech.edu
∗∗ Corresponding author. Department of Psychology, California National Primate Research Center, University of California, Davis, CA, 05616, USA. dfox@ucdavis.edu
1 These two authors contributed equally to this work.

07 4 2023
2023
07 4 2023
4 10008630 9 2022
5 2 2023
17 3 2023
© 2023 The Authors
2023
https://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Modern neuroscience approaches including optogenetics, calcium imaging, and other genetic manipulations have facilitated our ability to dissect specific circuits in rodent models to study their role in neurological disease. These approaches regularly use viral vectors to deliver genetic cargo (e.g., opsins) to specific tissues and genetically-engineered rodents to achieve cell-type specificity. However, the translatability of these rodent models, cross-species validation of identified targets, and translational efficacy of potential therapeutics in larger animal models like nonhuman primates remains difficult due to the lack of efficient primate viral vectors. A refined understanding of the nonhuman primate nervous system promises to deliver insights that can guide the development of treatments for neurological and neurodegenerative diseases. Here, we outline recent advances in the development of adeno-associated viral vectors for optimized use in nonhuman primates. These tools promise to help open new avenues for study in translational neuroscience and further our understanding of the primate brain.

Graphical abstract

Image 1

Highlights

• Nonhuman primates are a valuable model for study of human disease.

• Technical hurdles have impeded translation of gene delivery from mice to primates.

• Capsid engineering allows for systemic delivery to primate nervous system.

• Identification of gene regulatory elements may further enable cell-type specificity.

Keywords

Nonhuman primate
Adeno-associated viral vectors
Capsid variants
Enhancers
Brain
Gene delivery
==== Body
pmc1 Nonhuman primate as a valuable model for study of human diseases

A major goal of modern neuroscience is to inform our understanding of the human condition and brain-based disorders. However, this requires better comprehension of the distributed anatomical and molecularly diverse pathways, and functional circuits underlying disease. Recent advances in genetic technologies have made it possible to control and image neuronal circuits in living animals, through the delivery of various effectors, sensors, and reporters to the brain (Boyden et al., 2005; Fenno et al., 2011; Wang et al., 2019; Yang and Yuste, 2017). This breakthrough in technology has advanced our understanding of neural circuits, cell-types, molecules, neurotransmitters, and gene regulatory elements that work together to contribute to the progression of disease (e.g.(Coley et al., 2021; Cummings and Clem, 2020; Fadok et al., 2018; Pignatelli and Beyeler, 2019; Xu et al., 2019)). For example, research on anxiety-relevant circuits has leveraged optical control of specific cell-types (e.g. somatostatin and corticotrophin-releasing hormone expressing cells) and their projections to threat-relevant regions (e.g. central amygdala to periaqueductal gray interneurons) in order to elucidate multiple distinct mechanisms that underlie specific aspects of threat responding behavior (Ciocchi et al., 2010; Fadok et al., 2017; Holley and Fox, 2022; Tovote et al., 2016). This work has far-reaching implications for our understanding of anxiety disorders, by identifying multiple distinct mechanisms that likely contribute to differences in symptomatology. Similarly, in basic research studies of the mechanisms relevant to neurodegenerative diseases like Parkinson's, optical inhibition of cells in the subthalamic nucleus of parkinsonian rodents was sufficient to improve 6-hydroxydopamine-induced forelimb akinesia, opening the door to potential treatment avenues for patients with PD (Yoon et al., 2014). Unfortunately, these advances have not led to a large number of new and improved treatments for neurological diseases. This gap between rodent models of disease and translational outcomes is due, in part, to difficulty in validating the relevancy of these potential targets in human disease, as well as in understanding how these potential therapeutic compounds interact with their molecular targets in primates. Differences in evolutionary pressures have contributed to differences in brain structure and function across species, including the expansion of cortical regions during human evolution (Ahmed, 2018; Baker et al., 2020; Ménard et al., 2016; Pine et al., 2021; Preuss and Wise, 2022), potentially contributing to poor predictability of rodent disease models to the human condition. Nonhuman primates (NHPs) are well-suited to bridge this gap as their recent evolutionary divergence from a common ancestor have endowed them with many anatomical, physiological, and ethological similarities to humans (Kalin and Shelton, 2003; Öngür and Price, 2000; Petrides and Pandya, 1999, 2002; Phillips et al., 2014). This makes them well suited for anatomical and functional dissection in both the central nervous system (CNS) and peripheral nervous system (PNS), and as models for interrogation of potential therapeutic targets.

Perhaps the most notable distinction between human and rodent brains is the expansion of neocortex during human evolution (Kaas, 2012; Öngür and Price, 2000; Petrides and Pandya, 1999, 2002; Pine et al., 2021). This expansion is often thought to have contributed to the many high order abilities and social complexities related to human uniqueness (Kaas, 2012; Smaers et al., 2011). Many studies in humans have shed light on the neuronal circuits associated with these abilities, however, due to the limitations of the available tools for the study of the human brain, a more comprehensive understanding of the underlying biology is still needed (Craig, 2009; Cristofori et al., 2019; Gläscher et al., 2012; Horga et al., 2014). To this end, NHPs are of particular importance. To briefly review, NHPs can be roughly broken down into various simian species, which include monkeys and apes, and prosimians, such as lemurs. Monkeys can be further divided into Old World (Catarrhini) and New World (Platyrrhini) monkeys (Welker, 2017). Marmosets (Callithrix jacchus), which diverged from the human lineage approximately 35 million years ago (MYA), and rhesus macaques (Macaca mulatta), which diverged from humans even more recently, approximately ∼25 MYA (Fig. 1), are the two most common NHPs used in research. It is this recent evolutionary divergence from a common ancestor that has made NHPs a valuable model in neuroscience, as they possess a highly elaborated prefrontal cortex, including a well-developed internal granular layer (Bernardi and Salzman, 2019; Öngür and Price, 2000; Petrides and Pandya, 1999, 2002). Because they are our phylogenetic neighbors, NHPs share many behavioral and anatomical features with humans (Kalin and Shelton, 2003; Öngür and Price, 2000; Petrides and Pandya, 1999, 2002; Phillips et al., 2014). For example, the ability to navigate social complexities has been hypothesized to be enabled by the evolutionary expansion of the primate prefrontal cortex (Dunbar and Shultz, 2007; Pine et al., 2021). Indeed, unlike many animals, both NHPs and humans have developed complex social behaviors that have helped them navigate the complexities of living in large social groups (Chang and Platt, 2014). These include prosocial behaviors (Miller et al., 2016), social imitation (Subiaul et al., 2004), and in New World Monkeys like marmosets, monogamy and infant rearing (Miller et al., 2016; Saito, 2015). The shared social repertoires between monkeys and humans have been helpful in studying the underlying biology of social behaviors (Chang and Platt, 2014; Ziegler, 2018). In addition to social behaviors, the phylogenetic proximity of Old World monkeys, like rhesus macaques, to humans provides an avenue to study the primate brain which has a similar structure and cytoarchitecture to the human brain (Öngür and Price, 2000; Petrides and Pandya, 1999, 2002). For this reason, primates can contribute to understanding human-specific cognitive functions like higher-order cognition, attention, and working memory (Brady and Hampton, 2018; Deaner and Platt, 2003; Dezfouli et al., 2021; Rich and Wallis, 2016; Snyder et al., 2021; Xie et al., 2022).Fig. 1 Evolutionary tree depicting the phylogentic relationship of common research species. Among these species, Old World Monkeys, like the rhesus macaque, are approximately more similar to humans than New World Monkeys and rodents.

Fig. 1

While the phylogenetic proximity of NHPs to humans has made them anatomically and behaviorally similar, it is also likely that throughout evolution, the composition and function of neuronal circuits have adapted based on the evolutionary pressures placed on the species (Katz and Harris-Warrick, 1999). That is to say, while rodents and humans may share basic organization of circuits, changes within these circuits can cause large and important changes in behaviors (Katz and Harris-Warrick, 1999). Indeed, recent studies have found that while there seems to be a strong evolutionary conservation of cell-types between rodents and primates, there are also a number of primate-specific cell-types distributed throughout the brain-in cortical and subcortical structures and in midbrain (Hodge et al., 2019; Kamath et al., 2022; Krienen et al., 2020; Schmitz et al., 2022). For example, von Economo neurons and fork neurons which were originally assumed to exist only in humans and great apes, have been identified in Old World Monkeys (Evrard, 2019). This discovery provides the opportunity to elucidate the function of these neurons, as they have been shown to have projections to regions often implicated in studies of psychological dysfunction in both humans and NHPs (Craig, 2009; Evrard, 2019; Fox et al., 2015). Similarly, primates have a larger diversity of interneurons (Fishell and Kepecs, 2020; Franjic et al., 2022; Krienen et al., 2020; Schmitz et al., 2022), which may impact circuit dynamics in ways that cannot be studied in rodent models. For example, researchers have identified a primate-specific striatal interneuron (i.e. GABAergic neurons that express TAC3), which is thought to have emerged through a developmental repurposing of dopaminergic periglomerular cells of the olfactory bulb (Krienen et al., 2020; Schmitz et al., 2022). Additionally, a novel excitatory cell type expressing neuropeptide Y (NPY) was recently discovered in the primate visual cortex (Wei et al., 2022). As we aim to further dissect and elucidate the distributed brain-body circuits related to human pathology, the ability to target and manipulate specific cells will become increasingly important as they may be key determinants in the emergence of brain based disorders in humans.

In brief, rodent models have been useful in understanding the molecular and cellular basis of diseases, due in large part to advances in molecular genetic technologies (Boyden et al., 2005; Fenno et al., 2011; Yang and Yuste, 2017). However, rodents and humans differ in physiology, anatomy, and social complexity. To this end, NHPs are of particular importance because of the recent evolutionary divergence between humans and primates that have made them similar to humans both biologically and socially. However, a critical component of translation includes the translation of the tools that have been commonplace in rodent neuroscience. In this review, we highlight the need for more efficient neurotropic AAVs that can be delivered systemically in NHPs, recently engineered capsid variants that can cross the blood brain barrier in NHPs, and advances made to target specific cell-types.

2 AAVs enable gene delivery and circuit interrogation

The ability to define, monitor, and manipulate a neural circuit requires precise delivery of reporters, sensors, and effectors to the individual circuit components (e.g. cell-types). Viral vectors such as herpes simplex virus (HSV), rabies, adenovirus, lentivirus (LV), and adeno-associated viruses (AAVs) have emerged as an effective tool for neuroscience in that they enable neuronal tracing and functional interrogation through the delivery of various transgenes (Davidson and Breakefield, 2003; Ghosh et al., 2020; Hui et al., 2022; Kristensson et al., 1982; Liu et al., 2022). Viral vectors are composed of: i) a capsid, an outer protein shell enclosing the genetic material and which determines the vector's tropism, or ability to infect different cell-types; ii) regulatory elements such as enhancers or promoters which restrict expression to specific cell or tissue types; and iii) a transgene (Bulcha et al., 2021). Transgenes include fluorescent proteins as genetic reporters for visualization, sensors for measuring neurotransmitter release (e.g., GCaMP, DLight, etc.), opsins and synthetic receptors for cellular manipulation (e.g., ChR2, DREADDs), and repair templates for CRISPR-Cas9 based gene editing and expression manipulation (e.g., using CRISPRa/i) (Boyden et al., 2005; Klapoetke et al., 2014; X. Li et al., 2005; Magnus et al., 2011; Patriarchi et al., 2018; Roth, 2016; Yim et al., 2020; F. Zhang et al., 2007). Such genetic tools have advanced our understanding of how various cell-types and specific circuits contribute to adaptive behaviors and emergent properties of the brain. Among the viral vectors, AAVs are considered to be the safest since they are non-pathogenic, and are naturally replication deficient i.e. they lack the genes necessary for replication and replicate only when co-infected with a helper virus (Buller et al., 1981; Rose and Koczot, 1972). In contrast, lentiviruses transduce cells with higher efficiency than AAVs but there is uncertainty surrounding their safety due to the possibility of random insertional mutagenesis (Zheng et al., 2018). This can affect the genetic code at the DNA insertion site, leading to adverse outcomes, including cancer (Zheng et al., 2018). Similarly, herpes viral vectors can cause strong inflammatory responses (Ghosh et al., 2020). These non-specific and adverse effects have precluded them from widespread use in NHPs (Tremblay et al., 2020). Additionally, AAVs are capable of transducing both dividing and post-mitotic cells such as neurons, and so are ideally suited for gene manipulation studies that require stable, long term transgene expression in cells that have already matured (Bartlett et al., 1998). For these reasons, recombinant AAVs (rAAVs) have become the viral vector of choice for in vivo gene therapy applications, with more than 285 registered clinical trials to date (Kuzmin et al., 2021; U.S. National Library of Medicine (www.clinicaltrials.gov)).

For the study and treatment of neurological and neurodegenerative diseases, widespread distribution of transgene expression could be transformative (Hadaczek et al., 2016; Muramatsu, 2010; Sun and Roy, 2021). For example, idiopathic Parkinson's disease is hypothesized to result from the aggregation of a protein called alpha-synuclein (α-Syn) first in the enteric nervous system, before it propagates up the vagus nerve to the basal forebrain, midbrain and ultimately the cerebral cortex (Braak et al., 2003). rAAVs can be used to deliver a pathogenic protein such as α-Syn to model PD in animals, and help tease apart the cell-types in the ENS and CNS that are susceptible to α-Syn pathology (Alam et al., 2022; Challis et al., 2020; Huntington and Srinivasan, 2021; Kirik and Björklund, 2003; Ulusoy et al., 2010). In such cases, widespread transgene expression is required. Conversely, the pathogenic protein can be silenced, or the disease phenotype may be reversed by delivering a therapeutic gene such as GBA1, which encodes the lysosomal enzyme Glucerebrosidase, and has been shown to reduce inflammation and aggregation of α-Syn in models of PD as well as Gaucher's disease-a lysosomal neurodegenerative disorder (Abeliovich et al., 2021; Sardi et al., 2011, 2013). The route of AAV administration, dose, age at the time of injection, and preexisting neutralizing antibodies against it in the host, are all key determinants of an AAV's safety, efficacy and tropism (Chan and Deverman, 2022). There are many advantages to direct in-brain injection. In fact, most studies to date have directly injected viruses into the brain to deliver genetic cargo to specific regions. This has been performed in animal models of PD to target the putamen or substantia nigra (Bartus et al., 2013; Christine et al., 2009; Kaplitt et al., 2007; Kells et al., 2012; Muramatsu, 2010). Despite its many advantages, which include relatively dense and robust expression surrounding the infusion site, targeting large, diffuse, or spatially distributed regions can require multiple injections. Thus, this route of administration is most suitable for localized targets (Wang, 2021). Covering entire brain regions remains a challenge due to the size of the primate brain and often requires multiple craniotomies. Systemic delivery of AAVs obviates the need for multiple direct injections and importantly reduces the health risks associated with extremely long and highly invasive surgeries. As a therapeutic approach, systemic administration via a single injection might be a safer alternative toward achieving brain wide gene transduction (Bourdenx et al., 2014; Kimura and Harashima, 2022). Ultimately, increased efficacy of BBB-crossing AAVs may be combined with other technologies to achieve localized expression. However, crossing the BBB, which acts as a gatekeeper by preventing toxins and pathogens in the systemic circulation from entering the CNS, is a major hurdle for efficient gene delivery.

To date, 13 distinct naturally-occurring or wildtype serotypes of AAVs, (AAV1-13) have been identified in humans and NHPs (Srivastava, 2016). Each of these serotypes differs in capsid structure and, therefore, tropism (Agbandje-McKenna and Kleinschmidt, 2012). The most commonly used serotypes in rodent research are AAV2, AAV5, AAV8, and AAV9, which transduce the CNS, although some transduce other organs as well (Aschauer et al., 2013). In NHPs, the most commonly used serotypes are AAV5 and AAV9 (Tremblay et al., 2020). AAV9 has been particularly widely studied because of its ability to cross the BBB and has been employed in several CNS-targeted gene therapies (Chan and Deverman, 2022; Chen et al., 2021; Foust et al., 2009; Song et al., 2022; Yang et al., 2014; Zhang et al., 2011). Efforts have also been made to characterize other serotypes that also have the capability to cross the BBB. For example, Gao et al. cloned and identified more than 100 novel rAAVs from human and NHP tissues (Gao et al., 2005; Gao et al., 2002). Among these, AAVrh8, AAVrh10 and AAVhu32 were found to cross the BBB with high efficiencies, similar to AAV9.

However, these AAVs, including AAV9, have limitations that have prevented their wider use. For instance, their cell-type tropism can vary across species. In neonatal mice and macaques, intravenously administered AAV9 transduces neurons preferentially, whereas in juvenile and adult mice and macaques, the tropism shifts toward astrocytes (Bevan et al., 2011; Dehay et al., 2012; Foust et al., 2009; Gray et al., 2011; Mattar et al., 2013; Samaranch et al., 2012). Moreover, AAV9 and the other BBB crossing serotypes mentioned above have a higher tropism for peripheral organs such as the liver than the brain (Gray et al., 2011; Zincarelli et al., 2008). This is especially concerning in large animals such as NHPs as they require large volumes of virus for systemic delivery and the high doses of AAV needed to achieve clinical relevance can lead to hepatotoxicity or sensory neuron toxicity (Hinderer et al., 2018). Additionally, humans as well as NHPs harbor neutralizing anti-AAV antibodies to certain wildtype AAV serotypes from pre-existing exposure or develop anti-AAV antibodies after therapeutic rAAV administration (Louis Jeune et al., 2013). This is a limiting factor for gene therapy applications where subsequent viral administration may be needed if the transgene expression wanes over time. Another important consideration is the ∼4.7 kb size limitation of the AAV vector genome, which is comparatively smaller than that of lentiviruses (∼9.7 kb) or herpes simplex virus (∼150 kb) (Kumar et al., 2001; Latchman, 2001; Sena-Esteves et al., 2000).

When designing a study, it is important to take these considerations into account. In studies of the primate brain, it is important to ensure that the target gene sequence is reliably expressed, to minimize off-target effects, and to ensure animal safety. This has led most studies to prefer AAVs. However, if the genetic cargo is larger than optimal for an AAV genome, researchers run the risk of lower transduction efficiency, affecting their ability to perform the desired manipulation (Wu et al., 2010). These cost-benefit calculations are study-specific and constantly changing. Development of cell-type specificity, as a function of the viral capsid or shortened enhancer and promoter cargo, as described below could mitigate off-target effect. In the following sections, we will discuss current efforts to develop novel systemic rAAV vectors with high transduction efficiency and optimized biodistribution, with minimal off-target delivery, and low immunogenicity for gene delivery to the NHP CNS (Challis et al., 2022; Chan and Deverman, 2022; Davidson et al., 2022).

3 Capsid engineering for systemic delivery to achieve tissue specific biodistribution in NHPs

The capsid of an AAV is its primary point of interaction with receptors on the host cell surface which enable the virus to be internalized, and ultimately deliver their genetic cargo to the cell nucleus (Challis et al., 2022; Li and Samulski, 2020). Because of this, the capsid structure of AAVs have been widely researched in order to determine the protein domains responsible for cellular receptor binding, and consequently the virus' tropism and efficacy (Challis et al., 2022; Lee et al., 2018; Li and Samulski, 2020). Capsid modification or engineering is one route toward altering an AAV's tropism and efficacy as several permissive sites for rational and random amino acid substitutions and insertion have been identified (Challis et al., 2022). Through capsid engineering, we can enhance and refine AAV tropisms, as well as identify novel AAV serotypes with improved BBB crossing properties.

Capsid engineering can be carried out either through rational design or directed evolution. Rational design capitalizes on the knowledge of existing AAV serotypes to systematically predict and refine virus function (Lee et al., 2018). On the other hand, directed evolution is a high-throughput approach that involves using a selection process to generate variants with the desired properties such as antibody neutralization and/or tissue and cell-type tropism. By iteratively creating many non-naturally occurring viral serotypes and selecting those with the desired tropism for the next round of evaluation, researchers have been able to discover novel capsid variants that have a higher transduction efficiency at lower titers or concentration (Chan et al., 2017; Deverman et al., 2016; Körbelin et al., 2016; Kumar et al., 2020; Nonnenmacher et al., 2021). For example, our group has developed the Cre recombination-based AAV targeted evolution (CREATE) selection method (Deverman et al., 2016). In brief, the CREATE method enables the recovery of capsid sequences that transduce Cre-expressing cell populations in transgenic mice. Wherever Cre is present, a library fragment adjacent to the cap gene is inverted. PCR-based amplification can then detect the sequences that have successfully transduced the target population. After multiple rounds of evolution, top performing capsids can then be selected and characterized. This method led to the identification of AAV-PHP.B and AAV-PHP.eB, which target the mouse CNS (Chan et al., 2017; Deverman et al., 2016). Both AAV-PHP.B and AAV-PHP.eB are derived from AAV9, with AAV-PHP.eB showing >50% transduction of cells across brain regions of C57BL/6J mice (Chan et al., 2017). However, the ability of these engineered variants to transduce cells in the CNS of other mouse strains or NHPs is dependent on the administration route and whether it can cross the BBB (Mathiesen et al., 2020). Indeed, IV administration of AAV-PHP.B in marmosets showed poor transduction of neurons and astrocytes similar to AAV9 (Matsuzaki et al., 2018). Furthermore, Hordeaux et al. reported that a higher dose (7.9E13 gc/kg) of the AAV-PHP.B vector resulted in acute toxicity in an IV-injected macaque (Hordeaux et al., 2018). Subsequent studies revealed that the enhanced CNS tropism of AAV-PHP.B and AAV-PHP.eB in the C57BL/6J mice might be due to their interaction with the LY6A receptor, which is a GPI-anchored protein that is highly expressed by brain microvascular endothelial cells (Hordeaux et al., 2019; Mathiesen et al., 2020). Interestingly, there is no LY6A homolog in primates thus limiting the utility of these viruses in primates, and further highlighting the need for molecular target identification and validation studies in NHPs as a critical aspect of early research. We have begun to extend this directed evolution strategy to develop novel AAVs that can be used to target primate cells. Multiplexed-CREATE (M-CREATE) implements internal controls to reduce sequencing bias and increase the number of variants identified with enhanced CNS tropism (Kumar et al., 2020). This approach can be iterated across species, testing thousands of candidates in mice to identify top-performing capsids for primate testing. Recently, our lab used M-CREATE to identify systemic variants with enhanced CNS and PNS transduction in both Old World and New World monkeys. In marmosets, we found two variants evolved from AAV-PHP.eB, AAV-CAP-B10 and AAV-CAP-B22, to have enhanced CNS transduction after IV delivery in adult marmosets (Goertsen et al., 2022). AAV-CAP-B10 and AAV-CAP-B22 displayed four and two-fold increased neuronal transduction over AAV9, respectively. Furthermore, AAV-CAP-B10 showed decreased expression in the liver, where expression is typically associated with toxicity, as compared to AAV9. Importantly, broad and robust transgene expression was seen across cortical, subcortical, and cerebellar regions as well as in the dorsal root ganglia (DRG) and the spinal cord (SC).

More recently, we have developed AAV-MaCPNS1 and AAV-MaCPNS2 (Chen et al., 2022). Although these AAVs were designed to target the PNS in rodents, we found that they transduce PNS and CNS in both marmosets and rhesus macaques. Specifically, in adult marmosets, IV delivery of AAV-MaCPNS1/2 capsids carrying fluorescent reporter proteins (i.e., ssAAV:CAG-eGFP or ssAAV:CAG-tdTomato) targeted PNS and CNS more efficiently than AAV9. In the PNS, enhanced transduction was observed in DRG, the small intestine (SI), and the ascending fiber tracts in the dorsal column of the spinal cord (SC). Surprisingly, in the CNS, diffuse brain-wide transduction was seen in regions including the cortex, thalamus, globus pallidus, cerebellum, and brainstem. This tropism was recapitulated in infant rhesus macaques using IV delivery of AAV-MaCPNS1/2. In the PNS, enhanced transduction was seen in the SC, DRG, and gastrointestinal (GI) tract, including the esophagus, colon, and SI. Similar to what was observed in marmosets, in the rhesus monkey CNS, AAV-MaCPNS1/2 capsids mediated enhanced brain-wide transduction, including areas of the cortex, hippocampus, putamen, and brainstem. Additionally, AAV-MaCPNS1/2 displayed an increase in astrocyte transduction over AAV9 in the cortex and thalamus. However, of the two, MaCPNS2 had the greater fold increase in astrocytes. These results demonstrate the potential of these capsids in interrogating peripheral to brain circuitry.

In addition to M-CREATE, a multi species screening and characterization strategy was used to identify another capsid variant, AAV-CAP-Mac, for systemic brain-wide delivery in Old World Monkeys, including rhesus macaques and green monkeys (Chlorocebus sabaeus) (Chuapoco et al., in press). To characterize the transduction properties of AAV-CAP-Mac, pools of AAV-capsids, including AAV9, AAV-PHP.eB, AAV.CAP-B10 and other previously engineered AAVs, were simultaneously injected intravenously into infant rhesus macaques. Each capsid variant contained a single stranded human FXN (frataxin) transgene fused to a hemagglutinin (HA) epitope tag under control of a CAG promoter (ssCAG-hFXN-HA). AAV-CAP-Mac was found to primarily transduce primate neurons in all lobes of the cortex, cerebellum, and multiple subcortical regions. IV injection of AAV-CAP-Mac outperformed its parent capsid AAV9 as well as other previously engineered AAVs, including AAV-PHP.eB. Interestingly, AAV-CAP-Mac also outperformed AAV-CAP-B10, suggesting genetic differences in the BBB even between Old World and New World monkeys. AAV-CAP-Mac, was also shown to have reduced tropism towards the liver across primate species. Further validating the utility of AAV-CAP-Mac for NHP research, delivering a mixture of fluorescent proteins achieved a sparse Golgi stain-like expression pattern that could facilitate large-scale studies of neuronal morphology. Again, in this experiment, AAV-CAP-Mac was seen to target neurons in the cortex and multiple subcortical regions including hippocampus, putamen, thalamus, and caudate, enabling reconstruction of both medium spiny neurons and cortical pyramidal cells (Fig. 2). Additional characterization was also performed in 8-month-old green monkeys. Injections of AAV-CAP-Mac (packaging ssCAG-eGFP) via IV delivery showed broad and strong neuronal expression in all lobes of the cortex and in various subcortical regions. Consistent with results from the rhesus experiments, AAV-CAP-Mac transduced a higher percentage of neurons than AAV9 in sampled cortex and subcortical regions. In summary, these novel AAV variants with tailored properties and tropism, offer greater CNS and/or PNS transduction than AAV9 in NHP.Fig. 2 (A) Representative image of MaCPNS1-mediated expression of eGFP and MaCPNS2-mediated expression of tdTomato in the cerebellum and brainstem of an infant rhesus macaque. (B) Multicolor labeling of neurons in cortex after IV administration of a cocktail of 3 fluorescent proteins (ssCAG-mNeonGreen, ssCAG-mRuby2, and ssCAG-mTurquoise2) packaged in AAV-CAP-Mac and (C) morphological reconstruction of a striatal medium spiny neuron in rhesus macaque (scale, 20 uM). Scale bars for A and B = 100 uM.

Fig. 2

At present, in vivo characterization of AAV-CAP-Mac and AAV-MaCPNS1/2 has been limited to infant rhesus macaques (Chuapoco et al., in press). Higher vector doses could lead to increased immunogenicity due to preexisting neutralizing antibodies, thus, further characterization of these capsids in adult animals is necessary and currently underway. Encouragingly, though, these variants show significantly lower transduction of the liver compared to AAV9, potentially minimizing the risk of hepatotoxicity resulting from high systemic doses. Additionally, while these variants were engineered for systemic delivery, they may also be effective for local delivery. In vitro data from human-derived iPSCs supports the increased effectiveness of these AAVs over AAV9 (Chuapoco et al., in press), but further work is needed to test this, as well as the functional efficacy of the genetic cargo delivered by these novel AAVs.

In addition to our efforts, several other groups have also engineered AAVs with unique features that are relevant to NHPs. Using an in-vivo directed evolution strategy, Dalkara et al. engineered a novel AAV variant, 7m8, that specifically transduces photoreceptors in the NHP retina via intravitreal injections (Dalkara et al., 2013). Through iterative refinement, researchers have developed newly engineered variants that induce higher expression in foveal cones than 7m8 in cynomolgus macaques (Byrne et al., 2020). The in-vivo strategy DELIVER (directed evolution of AAV capsids leveraging in vivo expression of transgene RNA), has been used to identify muscle-tropic capsids that can be systemically administered to transduce primate muscles with high efficiency in cynomolgus macaques (Tabebordbar et al., 2021). Similarly, rAAV2-retro, which was developed for delivery of genetic cargo to retrogradely targeted cells, has also been tested for use in the primate brain (Bohlen et al., 2020) though see: (Cushnie et al., 2020)).

4 AAV-based approaches for targeting specific cell-types in NHPs

Developing tools to target specific cell-types to study their role in normal and disease circuitry remains a major challenge for primate neuroscience. Rodent models often rely on genetically-engineered Cre lines to achieve cell-type specificity. Unfortunately, primate gestational and maturational timelines preclude the widespread use of these genetic engineering approaches in primates (though see: (Drummer et al., 2021; Park et al., 2016; Sasaki et al., 2009; Tomioka et al., 2017)) so a different approach is needed. It is unlikely that capsid engineering alone will achieve the level of cell-type specificity required for NHP neuroscience research. This is due in large part because profiling AAV capsid variants generated by selection experiments is time-consuming and labor-intensive, and most remain uncharacterized (Zolotukhin and Vandenberghe, 2022). Although new molecular and computational tools, such as machine learning, might facilitate capsid profiling, these approaches also have limitations (Zolotukhin and Vandenberghe, 2022). Additionally, studies in NHPs suggest that novel transduction properties may not only arise from unique capsid binding properties, but also from uncharacterized capsid-promoter interactions (Bohlen et al., 2020). Therefore, it is likely that achieving cell-type specificity in NHPs will depend on a combination of BBB-crossing AAV capsid variants and regulatory elements.

Cis-acting regulatory DNA elements, such as promoters and enhancers, are sequences of DNA that proteins bind to in order to initiate and increase the likelihood of transcription respectively (Levine, 2010; Wittkopp and Kalay, 2012). Thus promoters and enhancers can determine the level of transgene expression and the cells they are expressed in. Ubiquitous promoters, such as cytomegalovirus (CMV), chicken β-actin (CBA), human elongation factor 1 alpha (EF1α) or combinations of these such as CMV early enhancer/chicken beta actin (CAG), drive high levels of transgene expression in most cell-types (Haery et al., 2019). However, high, widespread transgene expression is not always desired and can evoke immune responses to the transgene product (Perez et al., 2020; Samelson-Jones et al., 2020). Alternatively, cell-type specific promoters can be incorporated into the AAV cargo. These can be used, for instance, to target neurons (synapsin 1) or astrocytes (glial fibrillary acidic protein), or even more specifically dopaminergic neurons, cerebellar Purkinje cells, or parvalbumin (PVALB) neurons in the brain (El-Shamayleh et al., 2017; Hoshino et al., 2021; Matsuzaki et al., 2014; Nitta et al., 2017; Shinohara et al., 2016; Stauffer et al., 2016). Promoter sizes can range anywhere from ∼100 bp to 1000 bp (Domenger and Grimm, 2019). Due to the AAV's size limitation, ongoing efforts are focused on identifying shorter, and phylogenetically conserved, regulatory element sequences to direct cell-type specific transgene expression across species (de Leeuw et al., 2016; Domenger and Grimm, 2019; Matsuzaki et al., 2014; Nathanson, 2009).

Recently, chromatin profiling techniques coupled with next-generation sequencing led to the discovery of putative enhancers that are less than 600 bp and can drive cell-type specific activation of genes (Buenrostro et al., 2013; Cusanovich et al., 2015; Fang et al., 2021; Grandi et al., 2022; Graybuck et al., 2021; Hrvatin et al., 2019; Mich et al., 2021; Nair et al., 2020; Preissl et al., 2018; Rubin et al., 2020; Visel et al., 2013; Vormstein-Schneider et al., 2020). A distal-less homeobox (Dlx) gene enhancer sequence that targets GABAergic interneurons in the telencephalon of several vertebrate species including mouse and marmoset was identified (Dimidschstein et al., 2016; A. T. Lee et al., 2014; Zerucha et al., 2000). Additionally, the mouse ortholog of the Dlx5/6 enhancer (mDLX5/6), which is only ∼400 bp, packaged into either AAV1 or AAV9, showed similar specificity for GABAergic interneurons when locally injected into area V1 of the primary visual cortex of rhesus macaques (De et al., 2020). Mich et al. further optimized the human ortholog of the Dlx5/6 enhancer (hDLXI5/6i) by engineering a triple tandem of core elements taken from hDLXI5/6i and called it hDLX2.0 (Mich et al., 2021). AAV-PHP.eB containing hDLX2.0 upstream of a minimal beta-globin promoter and super yellow fluorescent protein-2 (SYFP2) reporter transduced GABAergic interneurons in ex vivo Macaca nemestrina cortical slice cultures and human neocortical slice cultures (Mich et al., 2021). Putative enhancers for targeting PVALB-expressing interneurons have similarly been identified, packaged in AAV-PHP.eB and tested in mice via retro-orbital injections, and in marmoset and macaque via local or intraparenchymal injections (Lawler et al., 2022; Mich et al., 2021; Vormstein-Schneider et al., 2020). These enhancers either targeted PVALB interneurons broadly or specific sub-classes of PVALB interneurons in the neocortex in both mouse and NHP. To identify regulatory elements that can drive faithful expression across species using AAV vectors, the selection method has largely focused on sequences that are conserved across species. However, Mich et al. reported that certain PVALB enhancer sequences present in the open chromatin analyses of the human neocortex but not in the mouse neocortex, were still able to drive selective expression in PVALB neurons in the mouse brain (Mich et al., 2021; Vormstein-Schneider et al., 2020). Thus, to minimize the number of experimental animals used for in vivo screening, it may be advantageous to develop machine-learning classifiers that can identify DNA sequence patterns important for driving species-agnostic cell-type specific activation (Lawler et al., 2022).

Single-cell and single-nucleus transcriptomics studies of the rodent and primate brain have revealed the molecular complexity and diversity of cell-types present based on their gene expression profiles (Hodge et al., 2019; Tasic et al., 2016, 2018; Zeisel et al., 2015). In the primary motor cortex alone, there are potentially 45 conserved cell-types among mouse, marmoset and human (Callaway et al., 2021). Only once a cell-type has been molecularly defined can researchers begin to identify DNA regulatory elements that are required for cell-type specific gene activation, and guide the development of tailored targeting strategies for treatment and functional interrogation. We have shown that broad CNS transduction in NHP is possible using the ubiquitous CAG promoter in our recently engineered vectors (AAV-CAP-Mac, AAV-MaCPNS1 and AAV-MaCPNS2) (Chen et al., 2022; Chuapoco et al., in press). These variants, which show comparatively higher neuronal transduction than AAV9 in NHPs, can be used to screen regulatory elements that specifically target neuronal subpopulations. One caveat of this approach is that injecting multiple AAVs with different enhancer and promoter elements in the same animal may cause interference between the regulatory elements, resulting in a loss of specificity compared to independent delivery (Mehta et al., 2019; Pouchelon et al., 2022). This may confound interpretation of pooled screens of putative regulatory elements.

Further cell- and tissue-type specificity can be achieved by incorporating microRNA (miRNA) target site sequences (∼22 nucleotides) into the 3’ untranslated coding region of AAVs. These bind to complementary miRNA sequences wherever they are expressed and inhibit mRNA expression. This approach has been used to detarget transgene expression from primary sensory neurons (miR183) and liver (miR122) to reduce dose-dependent toxicity in NHPs (Hordeaux et al., 2020; Kochunov et al., 2015). Recently, a database of miRNA expression across 196 primary cell-types was generated, enabling the potential testing of numerous combinations of miRNA-binding transgene cassettes (Patil et al., 2022). With spatial transcriptomics we can characterize the cell type-specific expression of various combinations of systemically-delivered AAV capsids and cargo, with the goal of expanding the gene delivery toolkit for NHPs (Jang et al., 2023).

5 Future directions

The effective delivery of sensors, effectors, and reporters for circuit tracing and manipulation, largely depends on the vector used. However, differences in brain size and immune function have hampered the widespread adoption of genetic technology in monkeys. Thus, the engineering of more efficient and specific viral vectors to target the CNS in NHPs addresses many of the challenges that have inhibited progress in translating rodent disease biology to better therapies and therapeutic approaches. Here, we review newly engineered systemic capsid variants that address these challenges. Specifically, using an adapted, cross-species directed evolution approach, our group has identified new variants that can transduce neuronal cells in CNS and PNS via peripheral injection in multiple NHP species commonly used in research.

In marmosets, AAV-CAP-B10 and AAV-CAP-B22, variants of AAV-PHP.eB, were identified to have enhanced CNS transduction compared to AAV9. In both marmosets and rhesus macaques, AAV-MaCPNS1 and AAV-MaCPNS2 variants transduced cells in both CNS and PNS. This may be particularly useful for studies where widespread transgene expression is desired as infusions in NHPs typically require multiple sites of injection to cover whole areas of tissue; however, this remains to be tested in relation to distributed brain function and/or behavior. In both rhesus macaques and green monkeys, AAV-CAP-Mac was found to transduce a higher percentage of neurons than AAV9. Importantly, these variants show significantly lower transduction in the liver compared to AAV9, thereby minimizing the risk of hepatotoxicity.

Still, while these novel AAVs may provide a new and necessary tool for delivering genetic cargo into the primate brain, many technical issues still remain. Below we briefly discuss these issues.1. Achieving cell-type specificity in NHPs. For decades, studies in NHPs have relied on lesions, reversible inactivation, and electrophysiology to elucidate the role of specific regions in a particular function (Balan et al., 2019; Dal Monte et al., 2015; Lak et al., 2014; Rudebeck et al., 2013). While this has provided invaluable insight into distributed circuits that underlie behavior, these studies are limited by their cell-type agnostic nature. Lesioning and reversible inactivation generally impact all cells in a particular region. In addition, lesion studies have resulted in conflicting reports on observed behaviors within the same region of the brain, often due to unintended damage to fibers of passage (Rudebeck et al., 2013). Similarly, electrophysiological recording techniques cannot differentiate molecular cell-types, and rely on electrophysiological-specific characterization (e.g. early-firing, late-firing, ramping, etc.). For example, in the ventral tegmental area, a minority of neurons share the same electrophysiological properties as dopamine neurons, leading to questions on whether some recordings have been misattributed to dopamine (Ungless and Grace, 2012). To address this issue, rodent studies have used TH-Cre mice to target dopamine-expressing VTA neurons for the expression of sensors and effectors (Bariselli et al., 2016; Lindeberg et al., 2004). However, in NHPs, Cre-lines do not yet exist or are not widely used. It is unlikely that capsid engineering alone will enable cell-type specificity. Instead, it is likely that a combination of capsid and DNA regulatory elements will achieve cell-type specificity. To this end, AAV-Cap-Mac and AAV-MaCPNS1/2 can be used to screen regulatory elements that specifically target neuronal subpopulations.

2. Effective delivery and functional efficacy of genetic cargo. Advances in genetic cargo, like opsins and DREADDs, have been critical in dissecting circuits that are thought to contribute to disease -- in rodents. However, these techniques have not been widely adopted in NHPs because of difficulties in delivering genetic cargo efficiently into the primate brain. Currently, only a few published studies in NHPs have demonstrated successful delivery of opsins, with AAV5 and AAV9 being amongst the most commonly used vectors (Tremblay et al., 2020). Moreover, outcome measures-anatomy, physiology, and behavior in these studies-vary greatly (Bliss-Moreau et al., 2022). For example, AAV5 has been shown to preferentially target some brain regions but not others (Roseboom et al., 2021). Still, a major hurdle remains in determining the extent in which systemic delivery can express effector-cargo in a sufficient proportion of cells to affect behavior. Even so, infecting a small proportion of cells can still help us better understand the contributions of a small number of cells on behaviors as the functional efficacy of sensor-cargo does not necessitate a large proportion of cells. Currently, the effective delivery and functional relevance of sensors and effectors have not been tested using AAV-CAP-Mac or AAV-MaCPNS1/2. Further work is needed to show the functional efficacy of the genetic cargo delivered by these novel AAVs.

Understanding the emergence or origins of brain-based disease requires coordinated cross-species research. To this end, NHP models are particularly important because of their shared biology with humans. However, tools for interrogating anatomical pathways and functional circuits will need to be translated for widespread use in NHPs. While the vectors presented here address many common technical challenges seen in NHPs, there is a continued need for more efficient and specific AAVs. We hope that further optimization of these vectors can lead to more efficient delivery and ultimately, lead to new tools for the study of the primate brain and the development of new treatments for brain-based disorders.

CRediT authorship contribution statement

Lillian J. Campos: Writing – original draft, Writing – review & editing. Cynthia M. Arokiaraj: Writing – original draft, Writing – review & editing. Miguel R. Chuapoco: Writing – review & editing. Xinhong Chen: Writing – review & editing. Nick Goeden: Writing – review & editing. Viviana Gradinaru: Writing – original draft, Writing – review & editing. Andrew S. Fox: Writing – original draft, Writing – review & editing.

Declaration of competing interest

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Viviana Gradinaru reports a relationship with Capsida Biotherapeutics that includes: board membership. Nick Goeden reports a relationship with Capsida Biotherapeutics that includes: employment.

Appendix A Supplementary data

The following is the Supplementary data to this article:Multimedia component 1

Multimedia component 1

Data availability

No data was used for the research described in the article.

Acknowledgments

Research in the Gradinaru and Fox labs is funded in part by Aligning Science Across Parkinson's [ASAP-020495] through the 10.13039/100000864 Michael J. Fox Foundation for Parkinson's Research (MJFF) (to V.G. and A.S.F.). For the purpose of open access, the author has applied a CC BY public copyright license to all Author Accepted Manuscripts arising from this submission. Additional support was provided by the 10.13039/100007862 California National Primate Research Center , NIH BRAIN Initiative Armamentarium grant UF1MH128336 (to V.G. and A.S.F.) and NIH P51OD011107 (to A.S.F). Figures were created using images from BioRender.com. We would like to thank Catherine Oikonomou and Rothem Kovner for help with manuscript editing.

Appendix A Supplementary data to this article can be found online at https://doi.org/10.1016/j.crneur.2023.100086.
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References

Abeliovich A. Hefti F. Sevigny J. Gene therapy for Parkinson's disease associated with GBA1 mutations J. Parkinsons Dis. 11 s2 2021 S183 S188 10.3233/JPD-212739 34151863
Agbandje-McKenna M. Kleinschmidt J. AAV capsid structure and cell interactions Snyder R. Moullier P. Adeno-Associated Virus: Methods and Protocols 807 2012 Humana Press 10.1007/978-1-61779-370-7
Ahmed S.H. Individual decision-making in the causal pathway to addiction: contributions and limitations of rodent models Pharmacol. Biochem. Behav. 164 2018 22 31 10.1016/j.pbb.2017.07.005 28709784
Alam M.M. Yang D. Li X.-Q. Liu J. Back T.C. Trivett A. Karim B. Barbut D. Zasloff M. Oppenheim J.J. Alpha synuclein, the culprit in Parkinson disease, is required for normal immune function Cell Rep. 38 2 2022 110090 10.1016/j.celrep.2021.110090
Aschauer D.F. Kreuz S. Rumpel S. Analysis of transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain PLoS One 8 9 2013 e76310 10.1371/journal.pone.0076310
Baker M. Hong S.-I. Kang S. Choi D.-S. Rodent models for psychiatric disorders: problems and promises Laboratory Animal Research 36 1 2020 9 10.1186/s42826-020-00039-z 32322555
Balan P.F. Gerits A. Zhu Q. Kolster H. Orban G.A. Wardak C. Vanduffel W. Fast compensatory functional Network changes caused by reversible inactivation of monkey parietal cortex Cerebr. Cortex 29 6 2019 2588 2606 10.1093/cercor/bhy128
Bariselli S. Glangetas C. Tzanoulinou S. Bellone C. Ventral tegmental area subcircuits process rewarding and aversive experiences J. Neurochem. 139 6 2016 1071 1080 10.1111/jnc.13779 27546491
Bartlett J.S. Samulski R.J. McCown T.J. Selective and rapid uptake of adeno-associated virus type 2 in brain Hum. Gene Ther. 9 8 1998 1181 1186 10.1089/hum.1998.9.8-1181 9625257
Bartus R.T. Baumann T.L. Siffert J. Herzog C.D. Alterman R. Boulis N. Turner D.A. Stacy M. Lang A.E. Lozano A.M. Olanow C.W. Safety/feasibility of targeting the substantia nigra with AAV2-neurturin in Parkinson patients Neurology 80 18 2013 1698 1701 10.1212/WNL.0b013e3182904faa 23576625
Bernardi S. Salzman C.D. The contribution of nonhuman primate research to the understanding of emotion and cognition and its clinical relevance Proc. Natl. Acad. Sci. USA 116 52 2019 26305 26312 10.1073/pnas.1902293116 31871162
Bevan A.K. Duque S. Foust K.D. Morales P.R. Braun L. Schmelzer L. Chan C.M. McCrate M. Chicoine L.G. Coley B.D. Porensky P.N. Kolb S.J. Mendell J.R. Burghes A.H. Kaspar B.K. Systemic gene delivery in large species for targeting spinal cord, brain, and peripheral tissues for pediatric disorders Mol. Ther. 19 11 2011 10.1038/mt.2011.157 Article 11
Bliss-Moreau E. Costa V.D. Baxter M.G. A pragmatic reevaluation of the efficacy of nonhuman primate optogenetics for psychiatry Oxford Open Neurosci. 1 2022 10.1093/oons/kvac006 kvac006
Bohlen M.O. McCown T.J. Powell S.K. El-Nahal H.G. Daw T. Basso M.A. Sommer M.A. Samulski R.J. Adeno-associated virus capsid-promoter interactions in the brain translate from rat to the Nonhuman primate Hum. Gene Ther. 31 21–22 2020 1155 1168 10.1089/hum.2020.196 32940068
Bourdenx M. Dutheil N. Bezard E. Dehay B. Systemic gene delivery to the central nervous system using Adeno-associated virus Front. Mol. Neurosci. 7 2014 10.3389/fnmol.2014.00050
Boyden E.S. Zhang F. Bamberg E. Nagel G. Deisseroth K. Millisecond-timescale, genetically targeted optical control of neural activity Nat. Neurosci. 8 9 2005 1263 1268 10.1038/nn1525 16116447
Braak H. R☐b U. Gai W.P. Del Tredici K. Idiopathic Parkinson's disease: possible routes by which vulnerable neuronal types may be subject to neuroinvasion by an unknown pathogen J. Neural. Transm. 110 5 2003 517 536 10.1007/s00702-002-0808-2 12721813
Brady R.J. Hampton R.R. Post-encoding control of working memory enhances processing of relevant information in rhesus monkeys (Macaca mulatta) Cognition 175 2018 26 35 10.1016/j.cognition.2018.02.012 29459237
Buenrostro J.D. Giresi P.G. Zaba L.C. Chang H.Y. Greenleaf W.J. Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position Nat. Methods 10 12 2013 1213 1218 10.1038/nmeth.2688 24097267
Bulcha J.T. Wang Y. Ma H. Tai P.W.L. Gao G. Viral vector platforms within the gene therapy landscape Signal Transduct. Targeted Ther. 6 1 2021 53 10.1038/s41392-021-00487-6
Buller R.M.L. Janik J.E. Sebring E.D. Rose J.A. Herpes simplex virus types 1 and 2 completely help adenovirus-associated virus replication J. Virol. 40 1 1981 241 247 10.1128/jvi.40.1.241-247.1981 6270377
Byrne L.C. Day T.P. Visel M. Strazzeri J.A. Fortuny C. Dalkara D. Merigan W.H. Schaffer D.V. Flannery J.G. In vivo–directed evolution of adeno-associated virus in the primate retina JCI Insight 5 10 2020 e135112 10.1172/jci.insight.135112
BRAIN Initiative Cell Census Network (BICCN), BRAIN Initiative Cell Census Network (BICCN) Corresponding authorsCallaway E.M. Dong H.-W. Ecker J.R. Hawrylycz M.J. Huang Z.J. Lein E.S. Ngai J. Osten P. Ren B. Tolias A.S. White O. Zeng H. Zhuang X. BICCN contributing principal investigatorsAscoli G.A. Behrens Chun M.M. J Sunkin S. A multimodal cell census and atlas of the mammalian primary motor cortex Nature 598 7879 2021 86 102 10.1038/s41586-021-03950-0 34616075
Challis C. Hori A. Sampson T.R. Yoo B.B. Challis R.C. Hamilton A.M. Mazmanian S.K. Volpicelli-Daley L.A. Gradinaru V. Gut-seeded α-synuclein fibrils promote gut dysfunction and brain pathology specifically in aged mice Nat. Neurosci. 23 3 2020 327 336 10.1038/s41593-020-0589-7 32066981
Challis R.C. Ravindra Kumar S. Chen X. Goertsen D. Coughlin G.M. Hori A.M. Chuapoco M.R. Otis T.S. Miles T.F. Gradinaru V. Adeno-associated virus toolkit to target diverse brain cells Annu. Rev. Neurosci. 45 1 2022 447 469 10.1146/annurev-neuro-111020-100834 35440143
Chan Y.A. Deverman B.E. Crossing the blood-brain barrier with AAVs: what's after SMA? de Lange E.C.M. Hammarlund-Udenaes M. Thorne R.G. Drug Delivery to the Brain 33 2022 Springer International Publishing 629 654 10.1007/978-3-030-88773-5_21
Chan K.Y. Jang M.J. Yoo B.B. Greenbaum A. Ravi N. Wu W.-L. Sánchez-Guardado L. Lois C. Mazmanian S.K. Deverman B.E. Gradinaru V. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems Nat. Neurosci. 20 8 2017 1172 1179 10.1038/nn.4593 28671695
Chang S.W.C. Platt M.L. Oxytocin and social cognition in rhesus macaques: implications for understanding and treating human psychopathology Brain Res. 1580 2014 57 68 10.1016/j.brainres.2013.11.006 24231551
Chen W. Yao S. Wan J. Tian Y. Huang L. Wang S. Akter F. Wu Y. Yao Y. Zhang X. BBB-crossing adeno-associated virus vector: an excellent gene delivery tool for CNS disease treatment J. Contr. Release 333 2021 129 138 10.1016/j.jconrel.2021.03.029
Chen X. Ravindra Kumar S. Adams C.D. Yang D. Wang T. Wolfe D.A. Arokiaraj C.M. Ngo V. Campos L.J. Griffiths J.A. Ichiki T. Mazmanian S.K. Osborne P.B. Keast J.R. Miller C.T. Fox A.S. Chiu I.M. Gradinaru V. Engineered AAVs for non-invasive gene delivery to rodent and non-human primate nervous systems Neuron 110 14 2022 10.1016/j.neuron.2022.05.003 Article 14
Christine C.W. Starr P.A. Larson P.S. Eberling J.L. Jagust W.J. Hawkins R.A. VanBrocklin H.F. Wright J.F. Bankiewicz K.S. Aminoff M.J. Safety and tolerability of putaminal AADC gene therapy for Parkinson disease Neurology 73 20 2009 1662 1669 10.1212/WNL.0b013e3181c29356 19828868
Chuapoco M.R. Flytzanis N.C. Goeden N. Octeau J.C. Roxas K.M. Chan K.Y. Scherrer J. Winchester J. Blackburn R.J. Campos L.J. Arokiaraj C.M. Miles T.F. Jang M.J. Vendemiatti J. Deverman B.E. Pickel J. Fox A.S. Gradinaru V. Intravenous gene transfer throughout the brain of infant old world primates using AAV Nat. Nanotechnol. 2023 n.d (in press)
Ciocchi S. Herry C. Grenier F. Wolff S.B.E. Letzkus J.J. Vlachos I. Ehrlich I. Sprengel R. Deisseroth K. Stadler M.B. Müller C. Lüthi A. Encoding of conditioned fear in central amygdala inhibitory circuits Nature 468 7321 2010 277 282 10.1038/nature09559 21068837
Coley A.A. Padilla-Coreano N. Patel R. Tye K.M. Valence processing in the PFC: reconciling circuit-level and systems-level views International Review of Neurobiology 158 2021 Elsevier 171 212 10.1016/bs.irn.2020.12.002
Craig A.D.B. How do you feel--now? The anterior insula and human awareness Nat. Rev. Neurosci. 10 1 2009 59 70 10.1038/nrn2555 19096369
Cristofori I. Cohen-Zimerman S. Grafman J. Executive functions Handbook of Clinical Neurology 163 2019 Elsevier 197 219 10.1016/B978-0-12-804281-6.00011-2
Cummings K.A. Clem R.L. Prefrontal somatostatin interneurons encode fear memory Nat. Neurosci. 23 1 2020 61 74 10.1038/s41593-019-0552-7 31844314
Cusanovich D.A. Daza R. Adey A. Pliner H.A. Christiansen L. Gunderson K.L. Steemers F.J. Trapnell C. Shendure J. Multiplex single-cell profiling of chromatin accessibility by combinatorial cellular indexing Science 348 6237 2015 910 914 10.1126/science.aab1601 25953818
Cushnie A.K. El-Nahal H.G. Bohlen M.O. May P.J. Basso M.A. Grimaldi P. Wang M.Z. de Velasco Ezequiel M.F. Sommer M.A. Heilbronner S.R. Using rAAV2-retro in rhesus macaques: promise and caveats for circuit manipulation J. Neurosci. Methods 345 2020 108859 10.1016/j.jneumeth.2020.108859
Dal Monte O. Costa V.D. Noble P.L. Murray E.A. Averbeck B.B. Amygdala lesions in rhesus macaques decrease attention to threat Nat. Commun. 6 1 2015 10161 10.1038/ncomms10161
Dalkara D. Byrne L.C. Klimczak R.R. Visel M. Yin L. Merigan W.H. Flannery J.G. Schaffer D.V. In vivo–directed evolution of a New adeno-associated virus for therapeutic outer retinal gene delivery from the vitreous Sci. Transl. Med. 5 189 2013 10.1126/scitranslmed.3005708
Davidson B.L. Breakefield X.O. Viral vectors for gene delivery to the nervous system Nat. Rev. Neurosci. 4 5 2003 353 364 10.1038/nrn1104 12728263
Davidson B.L. Gao G. Berry-Kravis E. Bradbury A.M. Bönnemann C. Buxbaum J.D. Corcoran G.R. Gray S.J. Gray-Edwards H. Kleiman R.J. Shaywitz A.J. Wang D. Zoghbi H.Y. Flotte T.R. Tauscher-Wisniewski S. Tifft C.J. Sahin M. Gene-based therapeutics for rare genetic neurodevelopmental psychiatric disorders Mol. Ther. 30 7 2022 2416 2428 10.1016/j.ymthe.2022.05.014 35585789
De A. El-Shamayleh Y. Horwitz G.D. Fast and reversible neural inactivation in macaque cortex by optogenetic stimulation of GABAergic neurons Elife 9 2020 e52658 10.7554/eLife.52658
de Leeuw C.N. Korecki A.J. Berry G.E. Hickmott J.W. Lam S.L. Lengyell T.C. Bonaguro R.J. Borretta L.J. Chopra V. Chou A.Y. D'Souza C.A. Kaspieva O. Laprise S. McInerny S.C. Portales-Casamar E. Swanson-Newman M.I. Wong K. Yang G.S. Zhou M. RAAV-compatible MiniPromoters for restricted expression in the brain and eye Mol. Brain 9 1 2016 52 10.1186/s13041-016-0232-4 27164903
Deaner R.O. Platt M.L. Reflexive social attention in monkeys and humans Curr. Biol. 13 18 2003 1609 1613 10.1016/j.cub.2003.08.025 13678591
Dehay B. Dalkara D. Dovero S. Li Q. Bezard E. Systemic scAAV9 variant mediates brain transduction in newborn rhesus macaques Sci. Rep. 2 1 2012 253 10.1038/srep00253 22355765
Deverman B.E. Pravdo P.L. Simpson B.P. Kumar S.R. Chan K.Y. Banerjee A. Wu W.-L. Yang B. Huber N. Pasca S.P. Gradinaru V. Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain Nat. Biotechnol. 34 2 2016 10.1038/nbt.3440 Article 2
Dezfouli M.P. Zarei M. Constantinidis C. Daliri M.R. Task-specific modulation of PFC activity for matching-rule governed decision-making Brain Struct. Funct. 226 2 2021 443 455 10.1007/s00429-020-02191-7 33398431
Dimidschstein J. Chen Q. Tremblay R. Rogers S.L. Saldi G.-A. Guo L. Xu Q. Liu R. Lu C. Chu J. Grimley J.S. Krostag A.-R. Kaykas A. Avery M.C. Rashid M.S. Baek M. Jacob A.L. Smith G.B. Wilson D.E. A viral strategy for targeting and manipulating interneurons across vertebrate species Nat. Neurosci. 19 12 2016 1743 1749 10.1038/nn.4430 27798629
Domenger C. Grimm D. Next-generation AAV vectors—do not judge a virus (only) by its cover Hum. Mol. Genet. 28 R1 2019 10.1093/hmg/ddz148 R3–R14
Drummer C. Vogt E.-J. Heistermann M. Roshani B. Becker T. Mätz-Rensing K. Kues W.A. Kügler S. Behr R. Generation and breeding of EGFP-transgenic marmoset monkeys: cell chimerism and implications for disease modeling Cells 10 3 2021 505 10.3390/cells10030505 33673402
Dunbar R.I.M. Shultz S. Evolution in the social brain Science 317 5843 2007 1344 1347 10.1126/science.1145463 17823343
El-Shamayleh Y. Kojima Y. Soetedjo R. Horwitz G.D. Selective optogenetic control of Purkinje cells in monkey cerebellum Neuron 95 1 2017 10.1016/j.neuron.2017.06.002 Article 1
Evrard H.C. The organization of the primate insular cortex Front. Neuroanat. 13 2019 43 10.3389/fnana.2019.00043 31133822
Fadok J.P. Krabbe S. Markovic M. Courtin J. Xu C. Massi L. Botta P. Bylund K. Müller C. Kovacevic A. Tovote P. Lüthi A. A competitive inhibitory circuit for selection of active and passive fear responses Nature 542 7639 2017 96 100 10.1038/nature21047 28117439
Fadok J.P. Markovic M. Tovote P. Lüthi A. New perspectives on central amygdala function Curr. Opin. Neurobiol. 49 2018 141 147 10.1016/j.conb.2018.02.009 29522976
Fang R. Preissl S. Li Y. Hou X. Lucero J. Wang X. Motamedi A. Shiau A.K. Zhou X. Xie F. Mukamel E.A. Zhang K. Zhang Y. Behrens M.M. Ecker J.R. Ren B. Comprehensive analysis of single cell ATAC-seq data with SnapATAC Nat. Commun. 12 1 2021 1337 10.1038/s41467-021-21583-9 33637727
Fenno L. Yizhar O. Deisseroth K. The development and application of optogenetics Annu. Rev. Neurosci. 34 1 2011 389 412 10.1146/annurev-neuro-061010-113817 21692661
Fishell G. Kepecs A. Interneuron types as attractors and controllers Annu. Rev. Neurosci. 43 1 2020 1 30 10.1146/annurev-neuro-070918-050421 31299170
Foust K.D. Nurre E. Montgomery C.L. Hernandez A. Chan C.M. Kaspar B.K. Intravascular AAV9 preferentially targets neonatal neurons and adult astrocytes Nat. Biotechnol. 27 1 2009 59 65 10.1038/nbt.1515 19098898
Fox A.S. Oler J.A. Shackman A.J. Shelton S.E. Raveendran M. McKay D.R. Converse A.K. Alexander A. Davidson R.J. Blangero J. Rogers J. Kalin N.H. Intergenerational neural mediators of early-life anxious temperament Proc. Natl. Acad. Sci. U. S. A 112 29 2015 9118 9122 10.1073/pnas.1508593112 26150480
Franjic D. Skarica M. Ma S. Arellano J.I. Tebbenkamp A.T.N. Choi J. Xu C. Li Q. Morozov Y.M. Andrijevic D. Vrselja Z. Spajic A. Santpere G. Li M. Zhang S. Liu Y. Spurrier J. Zhang L. Gudelj I. Transcriptomic taxonomy and neurogenic trajectories of adult human, macaque, and pig hippocampal and entorhinal cells Neuron 110 3 2022 452 469.e14 10.1016/j.neuron.2021.10.036 34798047
Gao G.-P. Alvira M.R. Wang L. Calcedo R. Johnston J. Wilson J.M. Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy Proc. Natl. Acad. Sci. USA 99 18 2002 11854 11859 10.1073/pnas.182412299 12192090
Gao G. Vandenberghe L. Wilson J. New recombinant serotypes of AAV vectors Curr. Gene Ther. 5 3 2005 285 297 10.2174/1566523054065057 15975006
Ghosh S. Brown A.M. Jenkins C. Campbell K. Viral vector systems for gene therapy: a comprehensive literature review of progress and biosafety challenges Appl. Biosafety 25 1 2020 7 18 10.1177/1535676019899502
Gläscher J. Adolphs R. Damasio H. Bechara A. Rudrauf D. Calamia M. Paul L.K. Tranel D. Lesion mapping of cognitive control and value-based decision making in the prefrontal cortex Proc. Natl. Acad. Sci. USA 109 36 2012 14681 14686 10.1073/pnas.1206608109 22908286
Goertsen D. Flytzanis N.C. Goeden N. Chuapoco M.R. Cummins A. Chen Y. Fan Y. Zhang Q. Sharma J. Duan Y. Wang L. Feng G. Chen Y. Ip N.Y. Pickel J. Gradinaru V. AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset Nat. Neurosci. 25 1 2022 106 115 10.1038/s41593-021-00969-4 34887588
Grandi F.C. Modi H. Kampman L. Corces M.R. Chromatin accessibility profiling by ATAC-seq Nat. Protoc. 17 6 2022 1518 1552 10.1038/s41596-022-00692-9 35478247
Gray S.J. Matagne V. Bachaboina L. Yadav S. Ojeda S.R. Samulski R.J. Preclinical differences of intravascular AAV9 delivery to Neurons and glia: a comparative study of adult mice and Nonhuman primates Mol. Ther. 19 6 2011 1058 1069 10.1038/mt.2011.72 21487395
Graybuck L.T. Daigle T.L. Sedeño-Cortés A.E. Walker M. Kalmbach B. Lenz G.H. Morin E. Nguyen T.N. Garren E. Bendrick J.L. Kim T.K. Zhou T. Mortrud M. Yao S. Siverts L.A. Larsen R. Gore B.B. Szelenyi E.R. Trader C. Enhancer viruses for combinatorial cell-subclass-specific labeling Neuron 109 9 2021 1449 1464.e13 10.1016/j.neuron.2021.03.011 33789083
Hadaczek P. Stanek L. Ciesielska A. Sudhakar V. Samaranch L. Pivirotto P. Bringas J. O'Riordan C. Mastis B. San Sebastian W. Forsayeth J. Cheng S.H. Bankiewicz K.S. Shihabuddin L.S. Widespread AAV1- and AAV2-mediated transgene expression in the nonhuman primate brain: implications for Huntington's disease Molecular Ther. Method. Clinic. Develop. 3 2016 16037 10.1038/mtm.2016.37
Haery L. Deverman B.E. Matho K.S. Cetin A. Woodard K. Cepko C. Guerin K.I. Rego M.A. Ersing I. Bachle S.M. Kamens J. Fan M. Adeno-associated virus technologies and methods for targeted Neuronal manipulation Front. Neuroanat. 13 2019 93 10.3389/fnana.2019.00093 31849618
Hinderer C. Katz N. Buza E.L. Dyer C. Goode T. Bell P. Richman L.K. Wilson J.M. Severe toxicity in Nonhuman primates and piglets following high-dose intravenous administration of an adeno-associated virus vector expressing human SMN Hum. Gene Ther. 29 3 2018 285 298 10.1089/hum.2018.015 29378426
Hodge R.D. Bakken T.E. Miller J.A. Smith K.A. Barkan E.R. Graybuck L.T. Close J.L. Long B. Johansen N. Penn O. Yao Z. Eggermont J. Höllt T. Levi B.P. Shehata S.I. Aevermann B. Beller A. Bertagnolli D. Brouner K. Conserved cell types with divergent features in human versus mouse cortex Nature 573 7772 2019 61 68 10.1038/s41586-019-1506-7 31435019
Holley D. Fox A.S. The central extended amygdala guides survival-relevant tradeoffs: implications for understanding common psychiatric disorders Neurosci. Biobehav. Rev. 142 2022 104879 10.1016/j.neubiorev.2022.104879
Hordeaux J. Wang Q. Katz N. Buza E.L. Bell P. Wilson J.M. The Neurotropic properties of AAV-PHP.B are limited to C57bl/6J mice Mol. Ther. 26 3 2018 10.1016/j.ymthe.2018.01.018 Article 3
Hordeaux J. Yuan Y. Clark P.M. Wang Q. Martino R.A. Sims J.J. Bell P. Raymond A. Stanford W.L. Wilson J.M. The GPI-linked protein LY6A drives AAV-PHP.B transport across the blood-brain barrier Mol. Ther. 27 5 2019 10.1016/j.ymthe.2019.02.013 Article 5
Hordeaux J. Buza E.L. Jeffrey B. Song C. Jahan T. Yuan Y. Zhu Y. Bell P. Li M. Chichester J.A. Calcedo R. Wilson J.M. MicroRNA-mediated inhibition of transgene expression reduces dorsal root ganglion toxicity by AAV vectors in primates Sci. Transl. Med. 12 569 2020 10.1126/scitranslmed.aba9188 Article 569
Horga G. Kaur T. Peterson B.S. Annual Research Review: current limitations and future directions in MRI studies of child- and adult-onset developmental psychopathologies JCPP (J. Child Psychol. Psychiatry) 55 6 2014 659 680 10.1111/jcpp.12185 24438507
Hoshino C. Konno A. Hosoi N. Kaneko R. Mukai R. Nakai J. Hirai H. GABAergic neuron-specific whole-brain transduction by AAV-PHP.B incorporated with a new GAD65 promoter Mol. Brain 14 1 2021 10.1186/s13041-021-00746-1 Article 1
Hrvatin S. Tzeng C.P. Nagy M.A. Stroud H. Koutsioumpa C. Wilcox O.F. Assad E.G. Green J. Harvey C.D. Griffith E.C. Greenberg M.E. A scalable platform for the development of cell-type-specific viral drivers Elife 8 2019 e48089 10.7554/eLife.48089
Hui Y. Zheng X. Zhang H. Li F. Yu G. Li J. Zhang J. Gong X. Guo G. Strategies for targeting Neural circuits: how to manipulate Neurons using virus vehicles Front. Neural Circ. 16 2022 882366 10.3389/fncir.2022.882366
Huntington T.E. Srinivasan R. Adeno-associated virus expression of α-synuclein as a tool to model Parkinson's disease: current understanding and knowledge gaps Aging and Disease 12 4 2021 1120 10.14336/AD.2021.0517 34221553
Jang M.J. Coughlin G.M. Jackson C.R. Chen X. Chuapoco M.R. Vendemiatti J.L. Wang A.Z. Gradinaru V. Spatial transcriptomics for profiling the tropism of viral vectors in tissues Nat. Biotechnol. 2023 10.1038/s41587-022-01648-w
Kaas J.H. The evolution of neocortex in primates Progress in Brain Research 195 2012 Elsevier 91 102 10.1016/B978-0-444-53860-4.00005-2
Kalin N.H. Shelton S.E. Nonhuman primate models to study anxiety, emotion regulation, and psychopathology Ann. N. Y. Acad. Sci. 1008 1 2003 189 200 10.1196/annals.1301.021 14998885
Kamath T. Abdulraouf A. Burris S.J. Langlieb J. Gazestani V. Nadaf N.M. Balderrama K. Vanderburg C. Macosko E.Z. Single-cell genomic profiling of human dopamine neurons identifies a population that selectively degenerates in Parkinson's disease Nat. Neurosci. 25 5 2022 588 595 10.1038/s41593-022-01061-1 35513515
Kaplitt M.G. Feigin A. Tang C. Fitzsimons H.L. Mattis P. Lawlor P.A. Bland R.J. Young D. Strybing K. Eidelberg D. During M.J. Safety and tolerability of gene therapy with an adeno-associated virus (AAV) borne GAD gene for Parkinson's disease: an open label, phase I trial Lancet 369 9579 2007 2097 2105 10.1016/S0140-6736(07)60982-9 17586305
Katz P.S. Harris-Warrick R.M. The evolution of neuronal circuits underlying species-specific behavior Curr. Opin. Neurobiol. 9 5 1999 628 633 10.1016/S0959-4388(99)00012-4 10508740
Kells A.P. Forsayeth J. Bankiewicz K.S. Glial-derived neurotrophic factor gene transfer for Parkinson's disease: anterograde distribution of AAV2 vectors in the primate brain Neurobiol. Dis. 48 2 2012 228 235 10.1016/j.nbd.2011.10.004 22019719
Kimura S. Harashima H. Non-invasive gene delivery across the blood-brain barrier: present and future perspectives Neural Regenerat. Res. 17 4 2022 785 10.4103/1673-5374.320981
Kirik D. Björklund A. Modeling CNS neurodegeneration by overexpression of disease-causing proteins using viral vectors Trends Neurosci. 26 7 2003 386 392 10.1016/S0166-2236(03)00164-4 12850435
Klapoetke N.C. Murata Y. Kim S.S. Pulver S.R. Birdsey-Benson A. Cho Y.K. Morimoto T.K. Chuong A.S. Carpenter E.J. Tian Z. Wang J. Xie Y. Yan Z. Zhang Y. Chow B.Y. Surek B. Melkonian M. Jayaraman V. Constantine-Paton M. Independent optical excitation of distinct neural populations Nat. Methods 11 3 2014 338 346 10.1038/nmeth.2836 24509633
Kochunov P. Jahanshad N. Marcus D. Winkler A. Sprooten E. Nichols T.E. Wright S.N. Hong L.E. Patel B. Behrens T. Jbabdi S. Andersson J. Lenglet C. Yacoub E. Moeller S. Auerbach E. Ugurbil K. Sotiropoulos S.N. Brouwer R.M. Heritability of fractional anisotropy in human white matter: a comparison of Human Connectome Project and ENIGMA-DTI data Neuroimage 111 2015 300 311 10.1016/j.neuroimage.2015.02.050 25747917
Körbelin J. Dogbevia G. Michelfelder S. Ridder D.A. Hunger A. Wenzel J. Seismann H. Lampe M. Bannach J. Pasparakis M. Kleinschmidt J.A. Schwaninger M. Trepel M. A brain microvasculature endothelial cell‐specific viral vector with the potential to treat neurovascular and neurological diseases EMBO Mol. Med. 8 6 2016 10.15252/emmm.201506078 Article 6
Krienen F.M. Goldman M. Zhang Q. del Rosario R C.H. Florio M. Machold R. Saunders A. Levandowski K. Zaniewski H. Schuman B. Wu C. Lutservitz A. Mullally C.D. Reed N. Bien E. Bortolin L. Fernandez-Otero M. Lin J.D. Wysoker A. Innovations present in the primate interneuron repertoire Nature 586 7828 2020 262 269 10.1038/s41586-020-2781-z 32999462
Kristensson K. Nennesmo I. Persson L. Lycke E. Neuron to neuron transmission of herpes simplex virus J. Neurol. Sci. 54 1 1982 149 156 10.1016/0022–510X(82)90227-1 6281393
Kumar M. Keller B. Makalou N. Sutton R.E. Systematic determination of the packaging limit of lentiviral vectors Hum. Gene Ther. 12 15 2001 1893 1905 10.1089/104303401753153947 11589831
Kumar S.R. Miles T.F. Chen X. Brown D. Dobreva T. Huang Q. Ding X. Luo Y. Einarsson P.H. Greenbaum A. Jang M.J. Deverman B.E. Gradinaru V. Multiplexed Cre-dependent selection yields systemic AAVs for targeting distinct brain cell types Nat. Methods 17 5 2020 541 550 10.1038/s41592-020-0799-7 32313222
Kuzmin D.A. Shutova M.V. Johnston N.R. Smith O.P. Fedorin V.V. Kukushkin Y.S. van der Loo J.C.M. Johnstone E.C. The clinical landscape for AAV gene therapies Nat. Rev. Drug Discov. 20 3 2021 173 174 10.1038/d41573-021-00017-7 33495615
Lak A. Stauffer W.R. Schultz W. Dopamine prediction error responses integrate subjective value from different reward dimensions Proc. Natl. Acad. Sci. USA 111 6 2014 2343 2348 10.1073/pnas.1321596111 24453218
Latchman D.S. Gene delivery and gene therapy with herpes simplex virus-based vectors Gene 264 1 2001 1 9 10.1016/S0378-1119(01)00322-5 11245972
Lawler A.J. Ramamurthy E. Brown A.R. Shin N. Kim Y. Toong N. Kaplow I.M. Wirthlin M. Zhang X. Phan B.N. Fox G.A. Wade K. He J. Ozturk B.E. Byrne L.C. Stauffer W.R. Fish K.N. Pfenning A.R. Machine learning sequence prioritization for cell type-specific enhancer design Elife 11 2022 e69571 10.7554/eLife.69571
Lee A.T. Gee S.M. Vogt D. Patel T. Rubenstein J.L. Sohal V.S. Pyramidal Neurons in prefrontal cortex receive subtype-specific forms of excitation and inhibition Neuron 81 1 2014 61 68 10.1016/j.neuron.2013.10.031 24361076
Lee E.J. Guenther C.M. Suh J. Adeno-associated virus (AAV) vectors: rational design strategies for capsid engineering Curr. Opinion Biomed. Eng. 7 2018 58 63 10.1016/j.cobme.2018.09.004
Levine M. Transcriptional enhancers in animal development and evolution Curr. Biol. 20 17 2010 R754 R763 10.1016/j.cub.2010.06.070 20833320
Li C. Samulski R.J. Engineering adeno-associated virus vectors for gene therapy Nat. Rev. Genet. 21 4 2020 255 272 10.1038/s41576-019-0205-4 32042148
Li X. Gutierrez D.V. Hanson M.G. Han J. Mark M.D. Chiel H. Hegemann P. Landmesser L.T. Herlitze S. Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopsin and green algae channelrhodopsin Proc. Natl. Acad. Sci. USA 102 49 2005 17816 17821 10.1073/pnas.0509030102 16306259
Lindeberg J. Usoskin D. Bengtsson H. Gustafsson A. Kylberg A. Söderström S. Ebendal T. Transgenic expression of Cre recombinase from the tyrosine hydroxylase locus: transgenic expression of Cre recombinase Genesis 40 2 2004 67 73 10.1002/gene.20065 15452869
Liu Q. Wu Y. Wang H. Jia F. Xu F. Viral tools for Neural circuit tracing Neurosci. Bull. 38 12 2022 1508 1518 10.1007/s12264-022-00949-z 36136267
Louis Jeune V. Joergensen J.A. Hajjar R.J. Weber T. Pre-existing anti–adeno-associated virus antibodies as a challenge in AAV gene therapy Hum. Gene Ther. Methods 24 2 2013 59 67 10.1089/hgtb.2012.243 23442094
Magnus C.J. Lee P.H. Atasoy D. Su H.H. Looger L.L. Sternson S.M. Chemical and genetic engineering of selective ion channel–ligand interactions Science 333 6047 2011 1292 1296 10.1126/science.1206606 21885782
Mathiesen S.N. Lock J.L. Schoderboeck L. Abraham W.C. Hughes S.M. CNS transduction benefits of AAV-PHP.eB over AAV9 are dependent on administration route and mouse strain Molecular Therapy. Methods & Clinical Development 19 2020 447 458 10.1016/j.omtm.2020.10.011 33294493
Matsuzaki Y. Oue M. Hirai H. Generation of a neurodegenerative disease mouse model using lentiviral vectors carrying an enhanced synapsin I promoter J. Neurosci. Methods 223 2014 133 143 10.1016/j.jneumeth.2013.12.004 24361760
Matsuzaki Y. Konno A. Mochizuki R. Shinohara Y. Nitta K. Okada Y. Hirai H. Intravenous administration of the adeno-associated virus-PHP.B capsid fails to upregulate transduction efficiency in the marmoset brain Neurosci. Lett. 665 2018 182 188 10.1016/j.neulet.2017.11.049 29175632
Mattar C.N. Waddington S.N. Biswas A. Johana N. Ng X.W. Fisk A.S. Fisk N.M. Tan L.G. Rahim A.A. Buckley S.M.K. Tan M.H. Lu J. Choolani M. Chan J.K.Y. Systemic delivery of scAAV9 in fetal macaques facilitates neuronal transduction of the central and peripheral nervous systems Gene Ther. 20 1 2013 69 83 10.1038/gt.2011.216 22278413
Mehta P. Kreeger L. Wylie D.C. Pattadkal J.J. Lusignan T. Davis M.J. Turi G.F. Li W.-K. Whitmire M.P. Chen Y. Kajs B.L. Seidemann E. Priebe N.J. Losonczy A. Zemelman B.V. Functional access to Neuron subclasses in rodent and primate forebrain Cell Rep. 26 10 2019 10.1016/j.celrep.2019.02.011 Article 10
Ménard C. Hodes G.E. Russo S.J. Pathogenesis of depression: insights from human and rodent studies Neuroscience 321 2016 138 162 10.1016/j.neuroscience.2015.05.053 26037806
Mich J.K. Graybuck L.T. Hess E.E. Mahoney J.T. Kojima Y. Ding Y. Somasundaram S. Miller J.A. Kalmbach B.E. Radaelli C. Gore B.B. Weed N. Omstead V. Bishaw Y. Shapovalova N.V. Martinez R.A. Fong O. Yao S. Mortrud M. Functional enhancer elements drive subclass-selective expression from mouse to primate neocortex Cell Rep. 34 13 2021 10.1016/j.celrep.2021.108754 Article 13
Miller C.T. Freiwald W.A. Leopold D.A. Mitchell J.F. Silva A.C. Wang X. Marmosets: a neuroscientific model of human social behavior Neuron 90 2 2016 219 233 10.1016/j.neuron.2016.03.018 27100195
Muramatsu S. The current status of gene therapy for Parkinson's disease Ann. Neurosci. 17 2 2010 10.5214/ans.0972-7531.1017209
Nair R.R. Blankvoort S. Lagartos M.J. Kentros C. Enhancer-Driven gene expression (EDGE) enables the generation of viral vectors specific to Neuronal subtypes iScience 23 3 2020 100888 10.1016/j.isci.2020.100888
Nathanson J.L. Short promoters in viral vectors drive selective expression in mammalian inhibitory neurons, but do not restrict activity to specific inhibitory cell-types Front. Neural Circ. 3 2009 10.3389/neuro.04.019.2009
Nitta K. Matsuzaki Y. Konno A. Hirai H. Minimal Purkinje cell-specific PCP2/L7 promoter virally available for rodents and Non-human primates Molecular Therapy. Methods & Clinical Development 6 2017 159 170 10.1016/j.omtm.2017.07.006 28828391
Nonnenmacher M. Wang W. Child M.A. Ren X.-Q. Huang C. Ren A.Z. Tocci J. Chen Q. Bittner K. Tyson K. Pande N. Chung C.H.-Y. Paul S.M. Hou J. Rapid evolution of blood-brain-barrier-penetrating AAV capsids by RNA-driven biopanning Molecular Ther. Method. Clinic. Develop. 20 2021 366 378 10.1016/j.omtm.2020.12.006
Öngür D. Price J.L. The organization of networks within the orbital and medial prefrontal cortex of rats, monkeys and humans Cerebr. Cortex 10 3 2000 206 219 10.1093/cercor/10.3.206
Park J.E. Zhang X.F. Choi S.-H. Okahara J. Sasaki E. Silva A.C. Generation of transgenic marmosets expressing genetically encoded calcium indicators Sci. Rep. 6 1 2016 34931 10.1038/srep34931
Patil A.H. Baran A. Brehm Z.P. McCall M.N. Halushka M.K. A Curated Human Cellular microRNAome Based on 196 Primary Cell Types 12 2022
Patriarchi T. Cho J.R. Merten K. Howe M.W. Marley A. Xiong W.-H. Folk R.W. Broussard G.J. Liang R. Jang M.J. Zhong H. Dombeck D. von Zastrow M. Nimmerjahn A. Gradinaru V. Williams J.T. Tian L. Ultrafast neuronal imaging of dopamine dynamics with designed genetically encoded sensors Science 360 6396 2018 eaat4422 10.1126/science.aat4422 29853555
Perez B.A. Shutterly A. Chan Y.K. Byrne B.J. Corti M. Management of neuroinflammatory responses to AAV-mediated gene therapies for Neurodegenerative diseases Brain Sci. 10 2 2020 119 10.3390/brainsci10020119 32098339
Petrides M. Pandya D.N. Dorsolateral prefrontal cortex: comparative cytoarchitectonic analysis in the human and the macaque brain and corticocortical connection patterns: dorsolateral prefrontal cortex in human and monkey Eur. J. Neurosci. 11 3 1999 1011 1036 10.1046/j.1460-9568.1999.00518.x 10103094
Petrides M. Pandya D.N. Comparative cytoarchitectonic analysis of the human and the macaque ventrolateral prefrontal cortex and corticocortical connection patterns in the monkey: ventrolateral prefrontal cortex in human and monkey Eur. J. Neurosci. 16 2 2002 291 310 10.1046/j.1460-9568.2001.02090.x 12169111
Phillips K.A. Bales K.L. Capitanio J.P. Conley A. Czoty P.W. ‘t Hart B.A. Hopkins W.D. Hu S.-L. Miller L.A. Nader M.A. Nathanielsz P.W. Rogers J. Shively C.A. Voytko M.L. Why primate models matter: why primate models matter Am. J. Primatol. 76 9 2014 801 827 10.1002/ajp.22281 24723482
Pignatelli M. Beyeler A. Valence coding in amygdala circuits Curr. Opinion Behavioral Sci. 26 2019 97 106 10.1016/j.cobeha.2018.10.010
Pine D.S. Wise S.P. Murray E.A. Evolution, emotion, and episodic engagement Am. J. Psychiatr. 178 8 2021 701 714 10.1176/appi.ajp.2020.20081187 34080889
Pouchelon G. Vergara J. McMahon J. Gorissen B.L. Lin J.D. Vormstein-Schneider D. Niehaus J.L. Burbridge T.J. Wester J.C. Sherer M. Fernandez-Otero M. Allaway K.C. Pelkey K. Chittajallu R. McBain C.J. Fan M. Nasse J.S. Wildenberg G.A. Fishell G. Dimidschstein J. A versatile viral toolkit for functional discovery in the nervous system Cell Rep. Method. 2 6 2022 100225 10.1016/j.crmeth.2022.100225
Preissl S. Fang R. Huang H. Zhao Y. Raviram R. Gorkin D.U. Zhang Y. Sos B.C. Afzal V. Dickel D.E. Kuan S. Visel A. Pennacchio L.A. Zhang K. Ren B. Single-nucleus analysis of accessible chromatin in developing mouse forebrain reveals cell-type-specific transcriptional regulation Nat. Neurosci. 21 3 2018 432 439 10.1038/s41593-018-0079-3 29434377
Preuss T.M. Wise S.P. Evolution of prefrontal cortex Neuropsychopharmacology 47 1 2022 3 19 10.1038/s41386-021-01076-5 34363014
Rich E.L. Wallis J.D. Decoding subjective decisions from orbitofrontal cortex Nat. Neurosci. 19 7 2016 973 980 10.1038/nn.4320 27273768
Rose J.A. Koczot F. Adenovirus-associated virus multiplication J. Virol. 10 1972
Roseboom P.H. Mueller S.A.L. Oler J.A. Fox A.S. Riedel M.K. Elam V.R. Olsen M.E. Gomez J.L. Boehm M.A. DiFilippo A.H. Christian B.T. Michaelides M. Kalin N.H. Evidence in primates supporting the use of chemogenetics for the treatment of human refractory neuropsychiatric disorders Mol. Ther. 29 12 2021 3484 3497 10.1016/j.ymthe.2021.04.021 33895327
Roth B.L. DREADDs for neuroscientists Neuron 89 4 2016 683 694 10.1016/j.neuron.2016.01.040 26889809
Rubin A.N. Malik R. Cho K.K.A. Lim K.J. Lindtner S. Robinson Schwartz S.E. Vogt D. Sohal V.S. Rubenstein J.L.R. Regulatory elements inserted into AAVs confer preferential activity in cortical interneurons Eneuro 7 6 2020 10.1523/ENEURO.0211-20.2020 ENEURO.0211-20.2020
Rudebeck P.H. Saunders R.C. Prescott A.T. Chau L.S. Murray E.A. Prefrontal mechanisms of behavioral flexibility, emotion regulation and value updating Nat. Neurosci. 16 8 2013 1140 1145 10.1038/nn.3440 23792944
Saito A. The marmoset as a model for the study of primate parental behavior Neurosci. Res. 93 2015 99 109 10.1016/j.neures.2014.12.011 25575642
Samaranch L. Salegio E.A. San Sebastian W. Kells A.P. Foust K.D. Bringas J.R. Lamarre C. Forsayeth J. Kaspar B.K. Bankiewicz K.S. Adeno-associated virus serotype 9 transduction in the central Nervous system of Nonhuman primates Hum. Gene Ther. 23 4 2012 382 389 10.1089/hum.2011.200 22201473
Samelson-Jones B.J. Finn J.D. Favaro P. Wright J.F. Arruda V.R. Timing of intensive immunosuppression impacts risk of transgene antibodies after AAV gene therapy in Nonhuman primates Molecular Ther. Method. Clinic. Develop. 17 2020 1129 1138 10.1016/j.omtm.2020.05.001
Sardi S.P. Clarke J. Kinnecom C. Tamsett T.J. Li L. Stanek L.M. Passini M.A. Grabowski G.A. Schlossmacher M.G. Sidman R.L. Cheng S.H. Shihabuddin L.S. CNS expression of glucocerebrosidase corrects α-synuclein pathology and memory in a mouse model of Gaucher-related synucleinopathy Proc. Natl. Acad. Sci. USA 108 29 2011 12101 12106 10.1073/pnas.1108197108 21730160
Sardi S.P. Clarke J. Viel C. Chan M. Tamsett T.J. Treleaven C.M. Bu J. Sweet L. Passini M.A. Dodge J.C. Yu W.H. Sidman R.L. Cheng S.H. Shihabuddin L.S. Augmenting CNS glucocerebrosidase activity as a therapeutic strategy for parkinsonism and other Gaucher-related synucleinopathies Proc. Natl. Acad. Sci. USA 110 9 2013 3537 3542 10.1073/pnas.1220464110 23297226
Sasaki E. Suemizu H. Shimada A. Hanazawa K. Oiwa R. Kamioka M. Tomioka I. Sotomaru Y. Hirakawa R. Eto T. Shiozawa S. Maeda T. Ito M. Ito R. Kito C. Yagihashi C. Kawai K. Miyoshi H. Tanioka Y. Generation of transgenic non-human primates with germline transmission Nature 459 7246 2009 523 527 10.1038/nature08090 19478777
Schmitz M.T. Sandoval K. Chen C.P. Mostajo-Radji M.A. Seeley W.W. Nowakowski T.J. Ye C.J. Paredes M.F. Pollen A.A. The development and evolution of inhibitory neurons in primate cerebrum Nature 603 7903 2022 871 877 10.1038/s41586-022-04510-w 35322231
Sena-Esteves M. Saeki Y. Fraefel C. Breakefield X.O. HSV-1 amplicon vectors—simplicity and versatility Mol. Ther. 2 1 2000 9 15 10.1006/mthe.2000.0096 10899823
Shinohara Y. Konno A. Takahashi N. Matsuzaki Y. Kishi S. Hirai H. Viral vector-based dissection of marmoset GFAP promoter in mouse and marmoset brains PLoS One 11 8 2016 e0162023 10.1371/journal.pone.0162023
Smaers J.B. Steele J. Case C.R. Cowper A. Amunts K. Zilles K. Primate prefrontal cortex evolution: human brains are the extreme of a lateralized ape trend Brain Behav. Evol. 77 2 2011 67 78 10.1159/000323671 21335939
Snyder A.C. Yu B.M. Smith M.A. A stable population code for attention in prefrontal cortex leads a dynamic attention code in visual cortex J. Neurosci. 41 44 2021 9163 9176 10.1523/JNEUROSCI.0608-21.2021 34583956
Song R. Pekrun K. Khan T.A. Zhang F. Paşca S.P. Kay M.A. Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell model Molecular Ther. Method. Clinic. Develop. 27 2022 73 88 10.1016/j.omtm.2022.09.002
Srivastava A. In vivo tissue-tropism of adeno-associated viral vectors Curr. Opin. Virol. 21 2016 75 80 10.1016/j.coviro.2016.08.003 27596608
Stauffer W.R. Lak A. Yang A. Borel M. Paulsen O. Boyden E.S. Schultz W. Dopamine Neuron-specific optogenetic stimulation in rhesus macaques Cell 166 6 2016 10.1016/j.cell.2016.08.024 Article 6
Subiaul F. Cantlon J.F. Holloway R.L. Terrace H.S. Cognitive Imitation in Rhesus Macaques 305 2004
Sun J. Roy S. Gene-based therapies for neurodegenerative diseases Nat. Neurosci. 24 3 2021 297 311 10.1038/s41593-020-00778-1 33526943
Tabebordbar M. Lagerborg K.A. Stanton A. King E.M. Ye S. Tellez L. Krunnfusz A. Tavakoli S. Widrick J.J. Messemer K.A. Troiano E.C. Moghadaszadeh B. Peacker B.L. Leacock K.A. Horwitz N. Beggs A.H. Wagers A.J. Sabeti P.C. Directed evolution of a family of AAV capsid variants enabling potent muscle-directed gene delivery across species Cell 184 19 2021 4919 4938.e22 10.1016/j.cell.2021.08.028 34506722
Tasic B. Menon V. Nguyen T.N. Kim T.K. Jarsky T. Yao Z. Levi B. Gray L.T. Sorensen S.A. Dolbeare T. Bertagnolli D. Goldy J. Shapovalova N. Parry S. Lee C. Smith K. Bernard A. Madisen L. Sunkin S.M. Adult mouse cortical cell taxonomy revealed by single cell transcriptomics Nat. Neurosci. 19 2 2016 335 346 10.1038/nn.4216 26727548
Tasic B. Yao Z. Graybuck L.T. Smith K.A. Nguyen T.N. Bertagnolli D. Goldy J. Garren E. Economo M.N. Viswanathan S. Penn O. Bakken T. Menon V. Miller J. Fong O. Hirokawa K.E. Lathia K. Rimorin C. Tieu M. Shared and distinct transcriptomic cell types across neocortical areas Nature 563 7729 2018 72 78 10.1038/s41586-018-0654-5 30382198
Tomioka I. Nogami N. Nakatani T. Owari K. Fujita N. Motohashi H. Takayama O. Takae K. Nagai Y. Seki K. Generation of transgenic marmosets using a tetracyclin-inducible transgene expression system as a neurodegenerative disease model Biol. Reprod. 97 5 2017 772 780 10.1093/biolre/iox129 29045563
Tovote P. Esposito M.S. Botta P. Chaudun F. Fadok J.P. Markovic M. Wolff S.B.E. Ramakrishnan C. Fenno L. Deisseroth K. Herry C. Arber S. Lüthi A. Midbrain circuits for defensive behaviour Nature 534 7606 2016 206 212 10.1038/nature17996 27279213
Tremblay S. Acker L. Afraz A. Albaugh D.L. Amita H. Andrei A.R. Angelucci A. Aschner A. Balan P.F. Basso M.A. Benvenuti G. Bohlen M.O. Caiola M.J. Calcedo R. Cavanaugh J. Chen Y. Chen S. Chernov M.M. Clark A.M. An open resource for Non-human primate optogenetics Neuron 108 6 2020 1075 1090.e6 10.1016/j.neuron.2020.09.027 33080229
Ulusoy A. Decressac M. Kirik D. Björklund A. Viral vector-mediated overexpression of α-synuclein as a progressive model of Parkinson's disease Progress in Brain Research 184 2010 Elsevier 89 111 10.1016/S0079-6123(10)84005-1
Ungless M.A. Grace A.A. Are you or aren't you? Challenges associated with physiologically identifying dopamine neurons Trends Neurosci. 35 7 2012 422 430 10.1016/j.tins.2012.02.003 22459161
U.S. National Library of Medicine. (n.d https://www.clinicaltrials.gov
Visel A. Taher L. Girgis H. May D. Golonzhka O. Hoch R.V. McKinsey G.L. Pattabiraman K. Silberberg S.N. Blow M.J. Hansen D.V. Nord A.S. Akiyama J.A. Holt A. Hosseini R. Phouanenavong S. Plajzer-Frick I. Shoukry M. Afzal V. A high-resolution enhancer atlas of the developing telencephalon Cell 152 4 2013 895 908 10.1016/j.cell.2012.12.041 23375746
Vormstein-Schneider D. Lin J.D. Pelkey K.A. Chittajallu R. Guo B. Arias-Garcia M.A. Allaway K. Sakopoulos S. Schneider G. Stevenson O. Vergara J. Sharma J. Zhang Q. Franken T.P. Smith J. Ibrahim L.A. Mastro K.J. Sabri E. Huang S. Viral manipulation of functionally distinct interneurons in mice, non-human primates and humans Nat. Neurosci. 23 12 2020 1629 1636 10.1038/s41593-020-0692-9 32807948
Wang D. Juggling safety and efficacy: finding ways to achieve both Hum. Gene Ther. 32 11–12 2021 538 539 10.1089/hum.2021.29164.dwa 34143660
Wang W. Kim C.K. Ting A.Y. Molecular tools for imaging and recording neuronal activity Nat. Chem. Biol. 15 2 2019 101 110 10.1038/s41589-018-0207-0 30659298
Wei J.-R. Hao Z.-Z. Xu C. Huang M. Tang L. Xu N. Liu R. Shen Y. Teichmann S.A. Miao Z. Liu S. Identification of visual cortex cell types and species differences using single-cell RNA sequencing Nat. Commun. 13 1 2022 6902 10.1038/s41467-022-34590-1 36371428
Welker B.J. The History of Our Tribe, Hominini 2017 Open SUNY Textbooks
Wittkopp P.J. Kalay G. Cis-regulatory elements: molecular mechanisms and evolutionary processes underlying divergence Nat. Rev. Genet. 13 1 2012 59 69 10.1038/nrg3095
Wu Z. Yang H. Colosi P. Effect of genome size on AAV vector packaging Mol. Ther. 18 1 2010 80 86 10.1038/mt.2009.255 19904234
Xie Y. Hu P. Li J. Chen J. Song W. Wang X.-J. Yang T. Dehaene S. Tang S. Min B. Wang L. Geometry of sequence working memory in macaque prefrontal cortex Science 375 6581 2022 632 639 10.1126/science.abm0204 35143322
Xu H. Liu L. Tian Y. Wang J. Li J. Zheng J. Zhao H. He M. Xu T.-L. Duan S. Xu H. A disinhibitory microcircuit mediates conditioned social fear in the prefrontal cortex Neuron 102 3 2019 668 682.e5 10.1016/j.neuron.2019.02.026 30898376
Yang W. Yuste R. In vivo imaging of neural activity Nat. Methods 14 4 2017 349 359 10.1038/nmeth.4230 28362436
Yang B. Li S. Wang H. Guo Y. Gessler D.J. Cao C. Su Q. Kramer J. Zhong L. Ahmed S.S. Zhang H. He R. Desrosiers R.C. Brown R. Xu Z. Gao G. Global CNS transduction of adult mice by intravenously delivered rAAVrh.8 and rAAVrh.10 and Nonhuman primates by rAAVrh.10 Mol. Ther. 22 7 2014 1299 1309 10.1038/mt.2014.68 24781136
Yim Y.Y. Teague C.D. Nestler E.J. In vivo locus-specific editing of the neuroepigenome Nat. Rev. Neurosci. 21 9 2020 471 484 10.1038/s41583-020-0334-y 32704051
Yoon H.H. Park J.H. Kim Y.H. Min J. Hwang E. Lee C.J. Francis Suh J.-K. Hwang O. Jeon S.R. Optogenetic inactivation of the subthalamic Nucleus improves forelimb akinesia in a rat model of Parkinson disease Neurosurgery 74 5 2014 533 541 10.1227/NEU.0000000000000297 24463495
Zeisel A. Muñoz-Manchado A.B. Codeluppi S. Lönnerberg P. La Manno G. Juréus A. Marques S. Munguba H. He L. Betsholtz C. Rolny C. Castelo-Branco G. Hjerling-Leffler J. Linnarsson S. Cell types in the mouse cortex and hippocampus revealed by single-cell RNA-seq Science 347 6226 2015 1138 1142 10.1126/science.aaa1934 25700174
Zerucha T. Stühmer T. Hatch G. Park B.K. Long Q. Yu G. Gambarotta A. Schultz J.R. Rubenstein J.L.R. Ekker M. A highly conserved enhancer in the dlx5/dlx6 intergenic region is the site of cross-regulatory interactions between Dlx genes in the embryonic forebrain J. Neurosci. 20 2 2000 709 721 10.1523/JNEUROSCI.20-02-00709.2000 10632600
Zhang F. Wang L.-P. Brauner M. Liewald J.F. Kay K. Watzke N. Wood P.G. Bamberg E. Nagel G. Gottschalk A. Deisseroth K. Multimodal fast optical interrogation of neural circuitry Nature 446 7136 2007 633 639 10.1038/nature05744 17410168
Zhang H. Yang B. Mu X. Ahmed S.S. Su Q. He R. Wang H. Mueller C. Sena-Esteves M. Brown R. Xu Z. Gao G. Several rAAV vectors efficiently cross the blood–brain barrier and transduce Neurons and astrocytes in the Neonatal mouse central Nervous system Mol. Ther. 19 8 2011 1440 1448 10.1038/mt.2011.98 21610699
Zheng C.-X. Wang S.-M. Bai Y.-H. Luo T.-T. Wang J.-Q. Dai C.-Q. Guo B.-L. Luo S.-C. Wang D.-H. Yang Y.-L. Wang Y.-Y. Lentiviral vectors and adeno-associated virus vectors: useful tools for gene transfer in pain research: lentiviral and adeno-associated virus vectors Anat. Rec. 301 5 2018 825 836 10.1002/ar.23723
Ziegler T.E. Measuring peripheral oxytocin and vasopressin in nonhuman primates Am. J. Primatol. 80 10 2018 e22871 10.1002/ajp.22871
Zincarelli C. Soltys S. Rengo G. Rabinowitz J.E. Analysis of AAV serotypes 1–9 mediated gene expression and tropism in mice after systemic injection Mol. Ther. 16 6 2008 1073 1080 10.1038/mt.2008.76 18414476
Zolotukhin S. Vandenberghe L.H. AAV capsid design: a Goldilocks challenge Trends Mol. Med. 28 3 2022 183 193 10.1016/j.molmed.2022.01.003 35093287