Biology

Prime editing with genuine Cas9 nickases minimizes unwanted indels

J. Lee, K. Lim, et al.

Discover how nCas9 nickases are changing the game in gene editing! This research by Jaesuk Lee and colleagues reveals the intricacies of nCas9 variants and their impact on DNA breaks and edits, paving the way for more precise genetic modifications.

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Playback language: English

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Abstract

Cas9 nickases (nCas9s), unlike CRISPR-Cas9 nucleases which create DNA double-strand breaks (DSBs), produce single-strand breaks. This study investigated off-target nicks using Digenome-seq. nCas9 (H840A), unlike nCas9 (D10A), created unwanted DSBs. Further mutagenesis in nCas9 (H840A), particularly the double-mutant nCas9 (H840A + N863A), eliminated DSB induction and reduced unwanted indels. In prime editor systems (PE2, PE3, ePE3), nCas9 (H840A + N854A) significantly increased correct edits while minimizing unwanted indels, achieving high editing purity.

Publisher

Nature Communications

Published On

Mar 30, 2023

Authors

Jaesuk Lee, Kayeong Lim, Annie Kim, Young Geun Mok, Eugene Chung, Sung-Ik Cho, Ji Min Lee, Jin-Soo Kim

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