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...

Descripción completa

Detalles Bibliográficos
Autores principales: Losito, Marialucrezia, Smith, Quentin M., Newton, Matthew D., Cuomo, Maria Emanuela, Rueda, David S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2021
Materias:
Chemistry
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8653695/
https://www.ncbi.nlm.nih.gov/pubmed/34494640
http://dx.doi.org/10.1039/d1cp03408a
_version_ 1784611718737952768
author Losito, Marialucrezia
Smith, Quentin M.
Newton, Matthew D.
Cuomo, Maria Emanuela
Rueda, David S.
author_facet Losito, Marialucrezia
Smith, Quentin M.
Newton, Matthew D.
Cuomo, Maria Emanuela
Rueda, David S.
author_sort Losito, Marialucrezia
collection PubMed
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.
format Online
Article
Text
id pubmed-8653695
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher The Royal Society of Chemistry
record_format MEDLINE/PubMed
spelling pubmed-86536952022-01-06 Cas12a target search and cleavage on force-stretched DNA Losito, Marialucrezia Smith, Quentin M. Newton, Matthew D. Cuomo, Maria Emanuela Rueda, David S. Phys Chem Chem Phys Chemistry 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. The Royal Society of Chemistry 2021-08-26 /pmc/articles/PMC8653695/ /pubmed/34494640 http://dx.doi.org/10.1039/d1cp03408a Text en This journal is © the Owner Societies https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Losito, Marialucrezia
Smith, Quentin M.
Newton, Matthew D.
Cuomo, Maria Emanuela
Rueda, David S.
Cas12a target search and cleavage on force-stretched DNA
title Cas12a target search and cleavage on force-stretched DNA
title_full Cas12a target search and cleavage on force-stretched DNA
title_fullStr Cas12a target search and cleavage on force-stretched DNA
title_full_unstemmed Cas12a target search and cleavage on force-stretched DNA
title_short Cas12a target search and cleavage on force-stretched DNA
title_sort cas12a target search and cleavage on force-stretched dna
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8653695/
https://www.ncbi.nlm.nih.gov/pubmed/34494640
http://dx.doi.org/10.1039/d1cp03408a
work_keys_str_mv AT lositomarialucrezia cas12atargetsearchandcleavageonforcestretcheddna
AT smithquentinm cas12atargetsearchandcleavageonforcestretcheddna
AT newtonmatthewd cas12atargetsearchandcleavageonforcestretcheddna
AT cuomomariaemanuela cas12atargetsearchandcleavageonforcestretcheddna
AT ruedadavids cas12atargetsearchandcleavageonforcestretcheddna

Ejemplares similares