Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a systemopen access
- 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
- Indexed
- SCIE
SCOPUS
- Journal Title
- MOLECULAR THERAPY-NUCLEIC ACIDS
- Volume
- 28
- Start Page
- 353
- End Page
- 362
- URI
- https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/170110
- 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 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.
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