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Increasing intracellular dNTP levels improves prime editing efficiency

  • Pengpeng Liu,
  • Karthikeyan Ponnienselvan,
  • Thomas Nyalile,
  • Sarah Oikemus,
  • Anya T. Joynt,
  • Sukanya Iyer,
  • Karen Kelly,
  • Dongsheng Guo,
  • Pyae P. Kyawe,
  • Emma Vanderleeden,
  • Sambra D. Redick,
  • Lei Huang,
  • Zexiang Chen,
  • Jeong Min Lee,
  • Celia A. Schiffer,
  • David M. Harlan,
  • Jennifer P. Wang,
  • Charles P. Emerson Jr,
  • Nathan D. Lawson,
  • Jonathan K. Watts,
  • Erik J. Sontheimer,
  • Jeremy Luban,
  • Scot A. Wolfe

摘要

In primary cell types, intracellular deoxynucleotide triphosphate (dNTP) levels are tightly regulated in a cell cycle-dependent manner. We report that prime editing efficiency is increased by mutations that improve the enzymatic properties of Moloney murine leukemia virus reverse transcriptase and treatments that increase intracellular dNTP levels. In combination, these modifications produce substantial increases in precise editing rates.