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Cas12a target search and cleavage on force-stretched DNA.


ABSTRACT: Using optical tweezers, we investigate target search and cleavage by CRISPR-Cas12a on force-stretched λ-DNA. Cas12a uses fast, one-dimensional hopping to locate its target. Binding and cleavage occur rapidly and specifically at low forces (≤5 pN), with a 1.8 nm rate-limiting conformational change. Mechanical distortion slows diffusion, increases off-target binding but hinders cleavage.

SUBMITTER: Losito M 

PROVIDER: S-EPMC8653695 | biostudies-literature |

REPOSITORIES: biostudies-literature

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