Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system

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Title
Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system
Author(s)
Hanseop Kim; Wi-Jae Lee; Chan Hyoung Kim; Yeounsun Oh; L W Gwon; H Lee; W Song; J K Hur; Kyung Seob LimKang Jin JeongKi Hoan NamYoung Suk WonKyeong-Ryoon LeeYoungjeon LeeYoung Hyun KimJae Won Huh; B H Jun; D S Lee; Seung Hwan Lee
Bibliographic Citation
Molecular Therapy-Nucleic Acids, vol. 28, pp. 353-362
Publication Year
2022
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.
ISSN
2162-2531
Publisher
Elsevier-Cell Press
DOI
http://dx.doi.org/10.1016/j.omtn.2022.03.021
Type
Article
Appears in Collections:
Ochang Branch Institute > Division of National Bio-Infrastructure > Futuristic Animal Resource & Research Center > 1. Journal Articles
Ochang Branch Institute > Division of National Bio-Infrastructure > National Primate Research Center > 1. Journal Articles
Ochang Branch Institute > Division of National Bio-Infrastructure > Laboratory Animal Resource & Research Center > 1. Journal Articles
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