A quantitative model for the dynamics of target recognition and off-target rejection by the CRISPR-Cas Cascade complex.

ABSTRACT: CRISPR-Cas effector complexes recognise nucleic acid targets by base pairing with their crRNA which enables easy re-programming of the target specificity in rapidly emerging genome engineering applications. However, undesired recognition of off-targets, that are only partially complementary to the crRNA, occurs frequently and represents a severe limitation of the technique. Off-targeting lacks comprehensive quantitative understanding and prediction. Here, we present a detailed analysis of the target recognition dynamics by the Cascade surveillance complex on a set of mismatched DNA targets using single-molecule supercoiling experiments. We demonstrate that the observed dynamics can be quantitatively modelled as a random walk over the length of the crRNA-DNA hybrid using a minimal set of pa

SUBMITTER: Rutkauskas M 

PROVIDER: S-EPMC9718816 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature