Targeted mutagenesis in mouse cells and embryos using an enhanced prime editor

Soo Ji Park, Tae Yeong Jeong, Seung Kyun Shin, Da Eun Yoon, Soo Yeon Lim, Sol Pin Kim, Jungmin Choi, Hyunji Lee, Jeong Im Hong, Jinhee Ahn, Je Kyung Seong, Kyoungmi Kim

Research output: Contribution to journalArticlepeer-review

Abstract

Prime editors, novel genome-editing tools consisting of a CRISPR-Cas9 nickase and an engineered reverse transcriptase, can induce targeted mutagenesis. Nevertheless, much effort is required to optimize and improve the efficiency of prime-editing. Herein, we introduce two strategies to improve the editing efficiency using proximal dead sgRNA and chromatin-modulating peptides. We used enhanced prime-editing to generate Igf2 mutant mice with editing frequencies of up to 47% and observed germline transmission, no off-target effects, and a dwarf phenotype. This improved prime-editing method can be efficiently applied to cell research and to generate mouse models.

Original languageEnglish
Article number170
JournalGenome Biology
Volume22
Issue number1
DOIs
Publication statusPublished - 2021 Dec

Keywords

  • Adamts20
  • Chromatin-modulating peptides
  • Dwarf phenotype
  • Germline transmission
  • Igf2
  • Mouse cells and embryos
  • Prime editor
  • Proximal dead sgRNA

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Genetics
  • Cell Biology

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