Cas12a target search and cleavage on force-stretched DNA.

Losito, Marialucrezia; Smith, Quentin M; Newton, Matthew D; Cuomo, Maria Emanuela; Rueda, David S

Losito, Marialucrezia; Smith, Quentin M; Newton, Matthew D; Cuomo, Maria Emanuela; Rueda, David S.

Afiliación

Losito M; Department of Infectious Disease, Section of Virology, Faculty of Medicine, Imperial College London, London W12 0NN, UK. david.rueda@imperial.ac.uk.
Smith QM; Single Molecule Imaging Group, MRC London Institute of Medical Sciences, London W12 0NN, UK.
Newton MD; Discovery Sciences, AstraZeneca, Cambridge CB4 0WG, UK.
Cuomo ME; Department of Infectious Disease, Section of Virology, Faculty of Medicine, Imperial College London, London W12 0NN, UK. david.rueda@imperial.ac.uk.
Rueda DS; Single Molecule Imaging Group, MRC London Institute of Medical Sciences, London W12 0NN, UK.

Phys Chem Chem Phys ; 23(47): 26640-26644, 2021 Dec 08.

Article en En | MEDLINE | ID: mdl-34494640

ABSTRACT

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.

Asunto(s)

Proteínas Bacterianas/química; Proteínas Asociadas a CRISPR/química; ADN/química; Endodesoxirribonucleasas/química; Sistemas CRISPR-Cas; Modelos Moleculares; Pinzas Ópticas

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Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Proteínas Bacterianas / ADN / Endodesoxirribonucleasas / Proteínas Asociadas a CRISPR Idioma: En Revista: Phys Chem Chem Phys Asunto de la revista: BIOFISICA / QUIMICA Año: 2021 Tipo del documento: Article País de afiliación: Reino Unido

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