Nucleotide metabolism constrains prime editing in hematopoietic stem and progenitor cells

ABSTRACT Therapeutic prime editing of hematopoietic stem and progenitor cells (HSPCs) holds great potential to remedy blood disorders. Since quiescent cells have low nucleotide levels and resist retroviral infection, we hypothesized that nucleotide metabolism could limit reverse transcription mediated prime editing in HSPCs. We demonstrate that deoxynucleoside supplementation and Vpx-mediated degradation of SAMHD1 improve prime editing efficiency in HSPCs, especially when coupled with editing approaches that evade mismatch repair.