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

作者：Soo-Ji Park, Tae Yeong Jeong, S. Shin, Da Eun Yoon, S. Lim, S. Kim, Jungmin Choi, Hyunji Lee, Jeong-Im Hong, Jinhee Ahn, Jeaseok Seong, Kyoungmi Kim · 发表于：Genome Biology · 年份：2021 · DOI：10.1186/s13059-021-02389-w · 被引用次数：6

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.