Cas12a target search and cleavage on force-stretched DNA

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.

Graphical abstract: Cas12a target search and cleavage on force-stretched DNA

Supplementary files

Article information

Article type
Communication
Submitted
26 Jul 2021
Accepted
26 Aug 2021
First published
26 Aug 2021
This article is Open Access
Creative Commons BY license

Phys. Chem. Chem. Phys., 2021, Advance Article

Cas12a target search and cleavage on force-stretched DNA

M. Losito, Q. M. Smith, M. D. Newton, M. E. Cuomo and D. S. Rueda, Phys. Chem. Chem. Phys., 2021, Advance Article , DOI: 10.1039/D1CP03408A

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements