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Res Sq
ResearchSquare
Research Square
2693-5015
American Journal Experts

39184105
10.21203/rs.3.rs-4863813/v1
10.21203/rs.3.rs-4863813
preprint
1
Article
PURE-seq identifies Egr1 as a Potential Master Regulator in Murine Aging by Sequencing Long-Term Hematopoietic Stem Cells
Abate Adam https://orcid.org/0000-0001-9614-4831

Pan Sixuan
Chang Kai-Chun https://orcid.org/0000-0001-7200-3532

Fernández-Maestre Inés
Haver Stéphane
Wereski Matthew
Bowman Robert
Levine Ross https://orcid.org/0000-0002-7884-1905

13 8 2024
rs.3.rs-4863813https://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-4863813/v1
nihpp-rs4863813v1.pdf
Abstract

Single-cell transcriptomics is valuable for uncovering individual cell properties, particularly in highly heterogeneous systems. However, this technique often results in the analysis of many well-characterized cells, increasing costs and diluting rare cell populations. To address this, we developed PURE-seq (PIP-seq for Rare-cell Enrichment and Sequencing) for scalable sequencing of rare cells. PURE-seq allows direct cell loading from FACS into PIP-seq reactions, minimizing handling and reducing cell loss. PURE-seq reliably captures rare cells, with 60 minutes of sorting capturing tens of cells at a rarity of 1 in 1,000,000. Using PURE-seq, we investigated murine long-term hematopoietic stem cells and their transcriptomes in the context of hematopoietic aging, identifying Egr1 as a potential master regulator of hematopoiesis in the aging context. PURE-seq offers an accessible and reliable method for isolating and sequencing cells that are currently too rare to capture successfully with existing methods.
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