Highly efficient genome editing in hematopoietic stem and progenitor cells

Achieving high knockout efficiencies in human hematopoietic stem progenitor cells (HSPCs) is of critical importance to study gene function and correlations. Here we have evaluated the most critical parameters for achieving highly efficient genome editing in HSPCs and make valuable recommendations. We demonstrate a fast and efficient method for gRNA selection and to genome edit HSPCs. We report knockout efficiencies up to 80% in human CD34+ HSPCs. Editing efficiency was similar between the different CD34+ progenitor and stem cell subpopulation including the immature CD34+CD38- subpopulation, which is enriched for hematopoietic stem cells. Ribonucleoprotein (RNP)-induced genome editing in human CD34+ HSPCs does not affect cell proliferation, differentiation or in vitro hematopoietic lineage commitment nor upregulate P21 expression. Overall, this research demonstrates that RNP editing of CD34+ HSPCs can be used for fast and efficient genome editing in human HSPCs paving the road for therapeutic implementation.