PRJNA529681
Targeted homology-directed repair in blood stem and progenitor cells with CRISPR nanoformulations
Ex vivo CRISPR gene editing in hematopoietic stem and progenitor cells has opened new treatment modalities for numerous diseases. The current process uses electroporation, sometimes followed by virus transduction. While this complex manipulation has resulted in high levels of gene editing at some genetic loci, cellular toxicity was observed. We have developed a CRISPR nanoformulation based on colloidal gold nanoparticles with a novel loading design capable of cellular entry without the need for electroporation or viruses. This highly monodispersed nanoformulation avoids lysosomal entrapment and localizes to the nucleus in primary human blood progenitors without toxicity. Nanoformulation-mediated gene editing is efficient and sustained with different CRISPR nucleases at multiple loci of therapeutic interest. Engraftment kinetics of nanoformulation-treated primary cells in humanized mice are better relative to non-treated cells, with no differences in differentiation. This is the first demonstration of nontoxic delivery of the entire CRISPR payload into primary human blood progenitors