Impact of CRISPR/HDR editing versus lentiviral transduction on long-term engraftment and clonal dynamics of HSPCs in rhesus macaques
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초록

For precise genome editing via CRISPR/homology-directed repair (HDR), effective and safe editing of long-term engrafting hematopoietic stem cells (LT-HSCs) is required. The impact of HDR on true LT-HSC clonal dynamics in a relevant large animal model has not been studied. To track the output and clonality of HDR-edited cells and to provide a comparison to lentivirally transduced HSCs in vivo, we developed a competitive rhesus macaque (RM) autologous transplantation model, co-infusing HSCs transduced with a barcoded GFP-expressing lentiviral vector (LV) and HDR edited at the CD33 locus. CRISPR/HDR-edited cells showed a two-log decrease by 2 months following transplantation, with little improvement via p53 inhibition, in comparison to minimal loss of LV-transduced cells long term. HDR long-term clonality was oligoclonal in contrast to highly polyclonal LV-transduced HSCs. These results suggest marked clinically relevant differences in the impact of current genetic modification approaches on HSCs. © 2024 Elsevier Inc.

키워드

CRISPR-Cas editingex vivogenetic barcodinghematopoietic stem cellshomology-directed repairHSC gene therapylentiviral gene additionprecise genome editingrhesus macaqueHEMATOPOIETIC STEM-CELLSHOMOLOGY-DIRECTED REPAIRPROGENITOR CELLSCRISPR-CAS9TRACKINGDNAINHIBITIONEFFICIENCY
제목
Impact of CRISPR/HDR editing versus lentiviral transduction on long-term engraftment and clonal dynamics of HSPCs in rhesus macaques
저자
Lee, Byung-ChulGin, AshleyWu, ChuanfengSingh, KomudiGrice, MaxMortlock, RylandAbraham, DianaFan, XingZhou, YifanAlJanahi, AishaChoi, UimookDeRavin, Suk SeeShin, TaehoonHong, SogunDunbar, Cynthia E.
DOI
10.1016/j.stem.2024.02.010
발행일
2024-04
유형
Article
저널명
Cell Stem Cell
31
4
페이지
455 ~ 466