Peptide-enabled ribonucleoprotein delivery for CRISPR engineering (PERC) in primary human immune cells and hematopoietic stem cells
Peptide-enabled ribonucleoprotein delivery for CRISPR engineering (PERC) is a new approach for genome editing of primary human cells. PERC uses a single amphiphilic peptide reagent to mediate intracellular delivery of the same pre-formed CRISPR ribonucleoprotein enzymes that are broadly employed in research and therapeutics, resulting in high-efficiency editing of immune cells and hematopoietic stem and progenitor cells (HSPCs). PERC facilitates nuclease-mediated gene knockout, precise transgene knock-in, and base editing. The protocol involves combination of the CRISPR ribonucleoprotein enzyme with peptide, followed by incubation with cultured cells. For efficient transgene knock-in, an adeno-associated virus (AAV) carrying homology-directed repair template DNA may be included.