A Genome Editing System for Therapeutical Targeting of Stem Cells.
Methods in molecular biology (Clifton, N.J.), 2185: 383-398 PMID 33165862 DOI 10.1007/978-1-0716-0810-4_24
Abstract
The CRISPR/Cas9 system can be exploited to disrupt genes or cis-regulatory elements in the genome of human hematopoietic stem cells. Here, we describe a protocol to deliver the CRISPR/Cas9 ribonucleoprotein complexes into primary human hematopoietic stem cells and to evaluate the engraftment and multilineage differentiation of edited cells in immunodeficient mice. This procedure allows the editing of a high proportion of long-term repopulating hematopoietic stem cells.