Cas12a target search and cleavage on force-stretched DNA

Cas12a target search and cleavage on force-stretched DNA

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. M...

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Veröffentlicht in:Physical chemistry chemical physics : PCCP 2021-12, Vol.23 (47), p.2664-26644
Hauptverfasser: Losito, Marialucrezia, Smith, Quentin M, Newton, Matthew D, Cuomo, Maria Emanuela, Rueda, David S
Format: Artikel
Sprache:eng
Schlagworte:
Bacterial Proteins - chemistry
Binding
Chemistry
Cleavage
CRISPR-Associated Proteins - chemistry
CRISPR-Cas Systems
Diffusion rate
DNA - chemistry
Endodeoxyribonucleases - chemistry
Models, Molecular
Optical Tweezers
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Zusammenfassung: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. CRISPR-Cas12a target search and cleavage on force-stretched λ -DNA using optical tweezers.
ISSN:1463-9076
1463-9084
DOI:10.1039/d1cp03408a