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

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 editin...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9035383/
https://www.ncbi.nlm.nih.gov/pubmed/35505967
http://dx.doi.org/10.1016/j.omtn.2022.03.021
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author 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
author_facet 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
author_sort Kim, Hanseop
collection PubMed
description 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.
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spelling pubmed-90353832022-05-02 Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system 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 Mol Ther Nucleic Acids Original Article 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. American Society of Gene & Cell Therapy 2022-03-28 /pmc/articles/PMC9035383/ /pubmed/35505967 http://dx.doi.org/10.1016/j.omtn.2022.03.021 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
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
Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system
title Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system
title_full Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system
title_fullStr Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system
title_full_unstemmed Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system
title_short Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system
title_sort highly specific chimeric dna-rna-guided genome editing with enhanced crispr-cas12a system
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9035383/
https://www.ncbi.nlm.nih.gov/pubmed/35505967
http://dx.doi.org/10.1016/j.omtn.2022.03.021
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