| Autor: |
Hanseop Kim, Wi-jae Lee, Chan Hyoung Kim, Yeounsun Oh, Lee Wha Gwon, Hyomin Lee, Woojeung Song, Junho K. Hur, Kyung-Seob Lim, Kang Jin Jeong, Ki-Hoan Nam, Young-Suk Won, Kyeong-Ryoon Lee, Youngjeon Lee, Young-Hyun Kim, Jae-Won Huh, Bong-Hyun Jun, Dong-Seok Lee, Seung Hwan Lee |
| Jazyk: |
angličtina |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
Molecular Therapy: Nucleic Acids, Vol 28, Iss , Pp 353-362 (2022) |
| Druh dokumentu: |
article |
| ISSN: |
2162-2531 |
| DOI: |
10.1016/j.omtn.2022.03.021 |
| Popis: |
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. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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