Addressing the dNTP bottleneck restricting prime editing activity

  • bioRxiv. 2023 Oct 29:2023.10.21.563443. doi: 10.1101/2023.10.21.563443.
Karthikeyan Ponnienselvan ,  Pengpeng Liu ,  Thomas Nyalile ,  Sarah Oikemus ,  Anya T Joynt ,  Karen Kelly ,  Dongsheng Guo ,  Zexiang Chen ,  Jeong Min Lee ,  Celia A Schiffer ,  Charles P Emerson ,  Nathan D Lawson ,  Jonathan K Watts ,  Erik J Sontheimer ,  Jeremy Luban ,  Scot A Wolfe
PMID: 37904991 DOI: 10.1101/2023.10.21.563443
Abstract

Prime editing efficiency is modest in cells that are quiescent or slowly proliferating where intracellular dNTP levels are tightly regulated. MMLV-reverse transcriptase - the prime editor polymerase subunit - requires high intracellular dNTPs levels for efficient polymerization. We report that prime editing efficiency in primary cells and in vivo is increased by mutations that enhance the enzymatic properties of MMLV-reverse transcriptase and can be further complemented by targeting SAMHD1 for degradation.

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