Cas12a target search and cleavage on force-stretched DNA

Phys Chem Chem Phys. 2021 Dec 8;23(47):26640-26644. doi: 10.1039/d1cp03408a.

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.

MeSH terms

  • Bacterial Proteins / chemistry*
  • CRISPR-Associated Proteins / chemistry*
  • CRISPR-Cas Systems
  • DNA / chemistry*
  • Endodeoxyribonucleases / chemistry*
  • Models, Molecular
  • Optical Tweezers

Substances

  • Bacterial Proteins
  • CRISPR-Associated Proteins
  • DNA
  • Cas12a protein
  • Endodeoxyribonucleases