Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system
- Authors
- Kim, Hanseop; Lee, Wi-jae; Kim, Chan Hyoung; Oh,Yeounsun; Gwon, Lee Wha; Lee, Hyomin; Song, Woojeung; Hur, Junho K.; Lim, Kyung-Seob; Jeong, Kang Jin; Nam,Ki-Hoan; Won,Young-Suk; Lee,Kyeong-Ryoon; Lee, Youngjeon; Kim,Young-Hyun; Huh, Jae-Won; Jun, Bong-Hyun; Lee, Dong-Seok; Lee, Seung Hwan
- Issue Date
- Jun-2022
- Publisher
- Cell Press
- Keywords
- chimeric DNA-RNA; efficient; en-Cas12a; gene therapy; genome editing; highly specific; RNA/DNA editing
- Citation
- Molecular Therapy - Nucleic Acids, v.28, pp 353 - 362
- Pages
- 10
- Journal Title
- Molecular Therapy - Nucleic Acids
- Volume
- 28
- Start Page
- 353
- End Page
- 362
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/61316
- DOI
- 10.1016/j.omtn.2022.03.021
- ISSN
- 2162-2531
- Abstract
- The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a system is composed of a Cas12a effector that acts as a DNA-cleaving endonuclease and a crispr RNA (crRNA) that guides the effector to the target DNA. It is considered a key molecule for inducing target-specific gene editing in various living systems. Here, we improved the efficiency and specificity of the CRISPR-Cas12a system through protein and crRNA engineering. In particular, to optimize the CRISPR-Cas12a system at the molecular level, we used a chimeric DNA-RNA guide chemically similar to crRNA to maximize target sequence specificity. Compared with the wild-type (wt)-Cas12a system, when using enhanced Cas12a system (en-Cas12a), the efficiency and target specificity improved on average by 2.58 and 2.77 times, respectively. In our study, when the chimeric DNA-RNA-guided en-Cas12a effector was used, the gene-editing efficiency and accuracy were simultaneously increased. These findings could contribute to highly accurate genome editing, such as human gene therapy, in the near future. © 2022 The Authors
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Collections - College of Natural Sciences > Department of Life Science > 1. Journal Articles
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