Expansion of the prime editing modality with Cas9 from Francisella novicida
Yeounsun Oh, Wi-jae Lee, Junho K. Hur, Woojeung Song, Youngjeon Lee, Hanseop Kim, Lee Wha Gwon, Younghyun 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
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.