Expansion of the prime editing modality with Cas9 from Francisella novicida

Abstract

Prime editing can induce a desired base substitution, insertion, or deletion in a target gene using reverse transcriptase (RT) after nick formation by CRISPR nickase. In this study, we developed 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 Cas 9 [FnCas9(H969A)] nickase module, which is a CRISPR-Cas9 ortholog. Using FnCas9(H969A) nickase, the targeting limitation of existing Streptococcus pyogenes Cas9 nickase [SpCas9(H840A)]-based prime editing was dramatically extended, and accurate prime editing was induced specifically for the target genes.

Expansion of the prime editing modality with Cas9 from Francisella novicida

“…Targeted deep sequencing data are available at NCBI Sequence Read Archive (SRA) under accession number PRJNA736246 [ 24 ]. The source code used for SNP analysis is released under an open source license (GNU General Public License V2.0) and available at https://github.com/yeounsunoh/Prime-editing-ClinVar-coverage-data.git [ 25 ].…”