Journal
CELL REPORTS
Volume 32, Issue 9, Pages -Publisher
CELL PRESS
DOI: 10.1016/j.celrep.2020.108093
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Funding
- Li Ka Shing Foundation
- Heritage Medical Research Institute
- NOMIS Foundation
- Lotte and Adolf Hotz-Sprenger Stiftung
- California Institute for Regenerative Medicine (CIRM) [TRAN1-09292]
- National Heart, Lung, and Blood Insitute of the National Institutes of Health (NIH) [DP2-HL-141006]
- National Institute on Aging of the NIH [T32 AG000266]
- Bill and Melinda Gates Foundation
- National Center of Competence in Research, RNA Disease
- Cancer Research Center (CRC) Zurich
- CIRM [TRAN1-09292]
- Siebel Stem Cell Center
- Swiss National Foundation [310030_184747/1]
- Clinical Research Priority Program ImmunoCure'' of the University of Zurich
- Swiss National Science Foundation (SNF) [310030_184747] Funding Source: Swiss National Science Foundation (SNF)
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Genome editing often takes the form of either error-prone sequence disruption by non-homologous end joining (NHEJ) or sequence replacement by homology-directed repair (HDR). Although NHEJ is generally effective, HDR is often difficult in primary cells. Here, we use a combination of immunophenotyping, next-generation sequencing, and single-cell RNA sequencing to investigate and reprogram genome editing outcomes in subpopulations of adult hematopoietic stem and progenitor cells. We find that although quiescent stem-enriched cells mostly use NHEJ, non-quiescent cells with the same immunophenotype use both NHEJ and HDR. Inducing quiescence before editing results in a loss of HDR in all cell subtypes. We develop a strategy of controlled cycling and quiescence that yields a 6-fold increase in the HDR/NHEJ ratio in quiescent stem cells ex vivo and in vivo. Our results highlight the tension between editing and cellular physiology and suggest strategies to manipulate quiescent cells for research and therapeutic genome editing.
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