한빛사논문
Byung-Chul Lee,1,2,3,8,* Ashley Gin,1,8 Chuanfeng Wu,1 Komudi Singh,1 Max Grice,1 Ryland Mortlock,1 Diana Abraham,1 Xing Fan,1 Yifan Zhou,1,4,5 Aisha AlJanahi,1 Uimook Choi,6 Suk See DeRavin,6 Taehoon Shin,1,7 Sogun Hong,1 and Cynthia E. Dunbar 1,9,*
1Translational Stem Cell Biology Branch, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA
2Department of Biological Sciences, Sookmyung Women’s University, Seoul, Korea
3Research Institute of Women’s Health, Sookmyung Women’s University, Seoul, Korea
4Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Puddicombe Way, Cambridge, UK
5Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge CB10 1SA, UK
6Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD, USA
7Department of Laboratory Animal Medicine, College of Veterinary Medicine, Jeju National University, Jeju, Korea
8These authors contributed equally
9Lead contact
*Corresponding author: correspondence to Byung-Chul Lee or Cynthia E. Dunbar
Abstract
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.
논문정보
관련 링크
연구자 키워드
관련분야 연구자보기
소속기관 논문보기
관련분야 논문보기
해당논문 저자보기