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 |
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| container_title | Physical chemistry chemical physics : PCCP |
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| creator | Losito, Marialucrezia Smith, Quentin M Newton, Matthew D Cuomo, Maria Emanuela Rueda, David S |
| description | 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. |
| doi_str_mv | 10.1039/d1cp03408a |
| format | Article |
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CRISPR-Cas12a target search and cleavage on force-stretched
λ
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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
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| fulltext | fulltext |
| identifier | ISSN: 1463-9076 |
| ispartof | Physical chemistry chemical physics : PCCP, 2021-12, Vol.23 (47), p.2664-26644 |
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| language | eng |
| recordid | cdi_crossref_primary_10_1039_D1CP03408A |
| source | MEDLINE; Royal Society Of Chemistry Journals 2008-; Alma/SFX Local Collection |
| subjects | Bacterial Proteins - chemistry Binding Chemistry Cleavage CRISPR-Associated Proteins - chemistry CRISPR-Cas Systems Diffusion rate DNA - chemistry Endodeoxyribonucleases - chemistry Models, Molecular Optical Tweezers |
| title | Cas12a target search and cleavage on force-stretched DNA |
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