Expansion of the prime editing modality with Cas9 from Francisella novicida

Oh, Y.; Lee, W.-j.; Kim, H.; Gwon, L. W.; Kim, Y.-H.; Park, Y.-H.; Kim, C. H.; Lim, K.-S.; Song, B.-S.; Huh, J.-W.; Kim, S.-U.; Jun, B.-H.; Jung, C.; Lee, S.

doi:10.1101/2021.05.25.445577

bioRxiv · 10.1101/2021.05.25.445577

Expansion of the prime editing modality with Cas9 from Francisella novicida

Oh, Y.·Lee, W.-j.·Kim, H.·Gwon, L. W.·Kim, Y.-H.·Park, Y.-H.·Kim, C. H.·Lim, K.-S.·Song, B.-S.·Huh, J.-W.·Kim, S.-U.·Jun, B.-H.·Jung, C.·Lee, S.

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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.

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Oh, Y., Lee, W.-j., Kim, H., Gwon, L. W., Kim, Y.-H., Park, Y.-H., Kim, C. H., Lim, K.-S., Song, B.-S., Huh, J.-W., Kim, S.-U., Jun, B.-H., Jung, C., Lee, S.. 2021-05-25. Expansion of the prime editing modality with Cas9 from Francisella novicida. https://doi.org/10.1101/2021.05.25.445577

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