Single-molecule perspectives of CRISPR/Cas systems: target search, recognition, and cleavage
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CRISPR/Cas systems have emerged as powerful tools for geneediting, nucleic acid detection, and therapeutic applications. Recent advances in single-molecule techniques have providednew insights into the DNA-targeting mechanisms of CRISPR/Cas systems, in particular, Types I, II, and V. Here, we reviewhow single-molecule approaches have expanded our understandingof key processes, namely target search, recognition,and cleavage. Furthermore, we focus on the dynamic behaviorof Cas proteins, including PAM site recognition and R-loopformation, which are crucial to ensure specificity and efficiencyin gene editing. Additionally, we discuss the conformationalchanges and interactions that drive precise DNA cleavage bydifferent Cas proteins. This mini review provides a comprehensiveoverview of CRISPR/Cas molecular dynamics, offeringconclusive insights into their broader potential for genomeediting and biotechnological applications.
Abstract
CRISPR/Cas systems have emerged as powerful tools for geneediting, nucleic acid detection, and therapeutic applications. Recent advances in single-molecule techniques have providednew insights into the DNA-targeting mechanisms of CRISPR/Cas systems, in particular, Types I, II, and V. Here, we reviewhow single-molecule approaches have expanded our understandingof key processes, namely target search, recognition,and cleavage. Furthermore, we focus on the dynamic behaviorof Cas proteins, including PAM site recognition and R-loopformation, which are crucial to ensure specificity and efficiencyin gene editing. Additionally, we discuss the conformationalchanges and interactions that drive precise DNA cleavage bydifferent Cas proteins. This mini review provides a comprehensiveoverview of CRISPR/Cas molecular dynamics, offeringconclusive insights into their broader potential for genomeediting and biotechnological applications.