Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9.

Research output: Contribution to journalArticle


Genome 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.


  • Pankaj K Mandal
  • Leonardo M R Ferreira
  • Ryan Collins
  • Torsten B Meissner
  • Christian L Boutwell
  • Max Friesen
  • Vladimir Vrbanac
  • Brian S Garrison
  • Alexei Stortchevoi
  • David Bryder
  • Kiran Musunuru
  • Harrison Brand
  • Andrew M Tager
  • Todd M Allen
  • Michael E Talkowski
  • Derrick J Rossi
  • Chad A Cowan
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Cell Biology
Original languageEnglish
Pages (from-to)643-652
JournalCell Stem Cell
Issue number5
Publication statusPublished - 2014
Publication categoryResearch

Bibliographic note

The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Stem Cell Aging (013212073)