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Res Sq
ResearchSquare
Research Square
2693-5015
American Journal Experts

10.21203/rs.3.rs-4810212/v1
10.21203/rs.3.rs-4810212
preprint
1
Article
Secreted Particle Information Transfer (SPIT) – A Cellular Platform for In Vivo Genetic Engineering
Nakauchi Hiromitsu https://orcid.org/0000-0002-9841-6973

Charlesworth Carsten
Homma Shota
Suchy Fabian https://orcid.org/0000-0002-7187-5360

Wang Sicong
Bhadhury Joydeep
Amaya Anais
Camarena Joab https://orcid.org/0000-0002-3102-5820

Zhang Jinyu
Tan Tze
Igarashi Kyomi
30 8 2024
rs.3.rs-4810212https://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License, which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
https://www.researchsquare.com/article/rs-4810212/v1
nihpp-rs4810212v1.pdf
Abstract

A multitude of tools now exist that allow us to precisely manipulate the human genome in a myriad of different ways. However, successful delivery of these tools to the cells of human patients remains a major barrier to their clinical implementation. Here we introduce a new cellular approach for in vivo genetic engineering, Secreted Particle Information Transfer (SPIT) that utilizes human cells as delivery vectors for in vivo genetic engineering. We demonstrate the application of SPIT for cell-cell delivery of Cre recombinase and CRISPR-Cas9 enzymes, we show that genetic logic can be incorporated into SPIT and present the first demonstration of human cells as a delivery platform for in vivo genetic engineering in immunocompetent mice. We successfully applied SPIT to genetically modify multiple organs and tissue stem cells in vivo including the liver, spleen, intestines, peripheral blood, and bone marrow. We anticipate that by harnessing the large packaging capacity of a human cell’s nucleus, the ability of human cells to engraft into patients’ long term and the capacity of human cells for complex genetic programming, that SPIT will become a paradigm shifting approach for in vivo genetic engineering.
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