CD34+ Cell RNP Nucleofection v1

Résumé

Human CD34+hematopoietic stem and progenitor cells (HSPCs) are a standard source of cells for clinical HSC transplantations as well as experimental xenotransplantation to generate “humanized mice”. To further extend the range of applications of these humanized mice, we developed a protocol to efficiently edit the genomes of human CD34+HSPCs before transplantation. We have optimized nucleofection of sgRNA:Cas9 ribonucleoprotein complexes and are now able to edit a candidate gene in CD34+HSPCs with almost 100% efficiency, and without affecting their potential for engraftment and multilineage differentiation in mice.

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