Cas12a target search and cleavage on force-stretched DNA
Marialucrezia Losito, Quentin M. Smith, Matthew D. Newton, Maria Emanuela Cuomo, David Rueda
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.
Topics & Concepts
Cleavage (geology)Optical tweezersCRISPRDNAMagnetic tweezersChemistryComputational biologyNanotechnologyPhysicsMaterials scienceBiologyOpticsBiochemistryComposite materialGeneFracture (geology)CRISPR and Genetic EngineeringAdvanced biosensing and bioanalysis techniquesRNA Interference and Gene Delivery