A cleavage rule for selection of increased-fidelity SpCas9 variants with high efficiency and no detectable off-targets.

ABSTRACT: Streptococcus pyogenes Cas9 (SpCas9) has been employed as a genome engineering tool with a promising potential within therapeutics. However, its off-target effects present major safety concerns for applications requiring high specificity. Approaches developed to date to mitigate this effect, including any of the increased-fidelity (i.e., high-fidelity) SpCas9 variants, only provide efficient editing on a relatively small fraction of targets without detectable off-targets. Upon addressing this problem, we reveal a rather unexpected cleavability ranking of target sequences, and a cleavage rule that governs the on-target and off-target cleavage of increased-fidelity SpCas9 variants but not that of SpCas9-NG or xCas9. According to this rule, for each target, an optimal variant with matching fi

SUBMITTER: Kulcsar PI 

PROVIDER: S-EPMC10505190 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature