Cas12a target search and cleavage on force-stretched DNA.
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ABSTRACT:
SUBMITTER: Losito M
PROVIDER: S-EPMC8653695 | biostudies-literature | 2021 Dec
REPOSITORIES: biostudies-literature
Publications
Cas12a target search and cleavage on force-stretched DNA.
Losito Marialucrezia M Smith Quentin M QM Newton Matthew D MD Cuomo Maria Emanuela ME Rueda David S DS
Physical chemistry chemical physics : PCCP 20211208 47
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. ...[more]
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