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

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

doi:10.1016/j.omtn.2022.03.021

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

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

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

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.

Original language	English
Pages (from-to)	353-362
Number of pages	10
Journal	Molecular Therapy Nucleic Acids
Volume	28
DOIs	https://doi.org/10.1016/j.omtn.2022.03.021
State	Published - 14 Jun 2022

Keywords

chimeric DNA-RNA
efficient
en-Cas12a
gene therapy
genome editing
highly specific
RNA/DNA editing

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10.1016/j.omtn.2022.03.021

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Kim, H., Lee, W. J., Kim, C. H., Oh, Y., Gwon, L. W., Lee, H., Song, W., Hur, J. K., Lim, K. S., Jeong, K. J., Nam, K. H., Won, Y. S., Lee, K. R., Lee, Y., Kim, Y. H., Huh, J. W., Jun, B. H., Lee, D. S., & Lee, S. H. (2022). Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system. Molecular Therapy Nucleic Acids, 28, 353-362. https://doi.org/10.1016/j.omtn.2022.03.021

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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.",

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author = "Hanseop Kim and Lee, {Wi jae} and Kim, {Chan Hyoung} and Yeounsun Oh and Gwon, {Lee Wha} and Hyomin Lee and Woojeung Song and Hur, {Junho K.} and Lim, {Kyung Seob} and Jeong, {Kang Jin} and Nam, {Ki Hoan} and Won, {Young Suk} and Lee, {Kyeong Ryoon} and Youngjeon Lee and Kim, {Young Hyun} and Huh, {Jae Won} and Jun, {Bong Hyun} and Lee, {Dong Seok} and Lee, {Seung Hwan}",

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AU - Kim, Chan Hyoung

AU - Oh, Yeounsun

AU - Gwon, Lee Wha

AU - Lee, Hyomin

AU - Song, Woojeung

AU - Hur, Junho K.

AU - Lim, Kyung Seob

AU - Jeong, Kang Jin

AU - Nam, Ki Hoan

AU - Won, Young Suk

AU - Lee, Kyeong Ryoon

AU - Lee, Youngjeon

AU - Kim, Young Hyun

AU - Huh, Jae Won

AU - Jun, Bong Hyun

AU - Lee, Dong Seok

AU - Lee, Seung Hwan

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Y1 - 2022/6/14

N2 - 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.

AB - 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.

KW - chimeric DNA-RNA

KW - efficient

KW - en-Cas12a

KW - gene therapy

KW - genome editing

KW - highly specific

KW - RNA/DNA editing

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EP - 362

JO - Molecular Therapy Nucleic Acids

JF - Molecular Therapy Nucleic Acids

ER -

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

Abstract

Keywords

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