Mandal, PankajFerreira, Leonardo Manuel RamosCollins, RyanMeissner, TorstenBoutwell, CFriesen, MaxVrbanac, VladimirGarrison, Brian ScottStortchevoi, AlexeiBryder, DavidMusunuru, KiranBrand, HarrisonTager, Andrew MartinAllen, ToddTalkowski, MichaelRossi, DerrickCowan, Chad2017-08-152014Mandal, Pankaj K., Leonardo M.R. Ferreira, Ryan Collins, Torsten B. Meissner, Christian L. Boutwell, Max Friesen, Vladimir Vrbanac, et al. 2014. “Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells Using CRISPR/Cas9.” In Cell Stem Cell 15, no. 5: 643–652. doi:10.1016/j.stem.2014.10.004.1934-5909http://nrs.harvard.edu/urn-3:HUL.InstRepos:33758654Genome editing via CRISPR/Cas9 has rapidly become the tool of choice by virtue of its efficacy and ease of use. However, CRISPR/Cas9-mediated genome editing in clinically relevant human somatic cells remains untested. Here, we report CRISPR/Cas9 targeting of two clinically relevant genes, B2M and CCR5, in primary human CD4+ T cells and CD34+ hematopoietic stem and progenitor cells (HSPCs). Use of single RNA guides led to highly efficient mutagenesis in HSPCs but not in T cells. A dual guide approach improved gene deletion efficacy in both cell types. HSPCs that had undergone genome editing with CRISPR/Cas9 retained multilineage potential. We examined predicted on- and off-target mutations via target capture sequencing in HSPCs and observed low levels of off-target mutagenesis at only one site. These results demonstrate that CRISPR/Cas9 can efficiently ablate genes in HSPCs with minimal off-target mutagenesis, which could have broad applicability for hematopoietic cell-based therapy.en-USJournal Article2015-07-29Mandal PK, Ferreira LM, Collins R, Meissner TB, Boutwell CL, Friesen M, Vrbanac V, Garrison BS, Stortchevoi A, Bryder D, Musunuru K, Brand H, Tager AM, Allen TM, Talkowski ME, Rossi DJ, Cowan CA2017-08-1510.1016/j.stem.2014.10.004