Engineered prime editors with PAM flexibility

Jiyeon Kweon, Jung Ki Yoon, An Hee Jang, Ha Rim Shin, Ji Eun See, Gayoung Jang, Jong Il Kim, Yongsub Kim

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Although prime editors are a powerful tool for genome editing, which can generate various types of mutations such as nucleotide substitutions, insertions, and deletions in the genome without double-strand breaks or donor DNA, the conventional prime editors are still limited to their target scopes because of the PAM preference of the Streptococcus pyogenes Cas9 (spCas9) protein. Here, we describe the engineered prime editors to expand the range of their target sites using various PAM-flexible Cas9 variants. Using the engineered prime editors, we could successfully generate more than 50 types of mutations with up to 51.7% prime-editing activity in HEK293T cells. In addition, we successfully introduced the BRAF V600E mutation, which could not be induced by conventional prime editors. These variants of prime editors will broaden the applicability of CRISPR-based prime editing technologies in biological research.

Original languageEnglish
Pages (from-to)2001-2007
Number of pages7
JournalMolecular Therapy
Volume29
Issue number6
DOIs
StatePublished - 2 Jun 2021
Externally publishedYes

Keywords

  • CRISPR-Cas
  • PAM variants
  • genome editing
  • prime editing

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