Expansion of the prime editing modality with Cas9 from Francisella novicida

Yeounsun Oh, Wi jae Lee, Junho K. Hur, Woo Jeung Song, Youngjeon Lee, Hanseop Kim, Lee Wha Gwon, Young Hyun Kim, Young Ho Park, Chan Hyoung Kim, Kyung Seob Lim, Bong Seok Song, Jae Won Huh, Sun Uk Kim, Bong Hyun Jun, Cheulhee Jung, Seung Hwan Lee

Research output: Contribution to journalArticlepeer-review

Abstract

Prime editing can induce a desired base substitution, insertion, or deletion in a target gene using reverse transcriptase after nick formation by CRISPR nickase. In this study, we develop a technology that can be used to insert or replace external bases in the target DNA sequence by linking reverse transcriptase to the Francisella novicida Cas9, which is a CRISPR-Cas9 ortholog. Using FnCas9(H969A) nickase, the targeting limitation of existing Streptococcus pyogenes Cas9 nickase [SpCas9(H840A)]-based prime editing is dramatically extended, and accurate prime editing is induced specifically for the target genes in human cell lines.

Original languageEnglish
Article number92
JournalGenome Biology
Volume23
Issue number1
DOIs
Publication statusPublished - 2022 Dec

Keywords

  • CRISPR-Cas9
  • Francisella novicida
  • Ortholog
  • Prime editing
  • Target expansion

ASJC Scopus subject areas

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

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