pISSN 2288-6982
eISSN 2288-7105



Mini review

BioDesign 2023; 11(1): 11-15

Published online March 30, 2023

© Korean Society for Structural Biology

Single-molecule fluorescence assays for studying the mechanism of DNA cleavage by CRISPR-Cas12a

Yuyoung Kim and Sanghwa Lee*

Department of Medical Life Sciences and Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea

Correspondence to: *

Received: February 28, 2023; Revised: March 24, 2023; Accepted: March 24, 2023


CRISPR-Cas12a system has been widely utilized for genome editing and molecular diagnostics applications. However, existing CRISPR-Cas12a systems still have much room for improving its functionality to expand their applications. To achieve this goal, a thorough understanding of the conformational dynamics of CRISPR-Cas12a system is essentially required. In this regard, single-molecule fluorescence assays have been recently applied to study the dynamics of CRISPR-Cas12a. In this review, we introduce single-molecule fluorescence studies on CRISPR-Cas12a system and provide practical guidelines required for reliable implementation of the single-molecule fluorescence methods to study the molecular mechanism of DNA cleavage reaction of CRISPR-Cas12a system.