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A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing

The CRISPR-Cas system can treat autosomal dominant diseases by nonhomologous end joining (NHEJ) gene disruption of mutant alleles. However, many single-nucleotide mutations cannot be discriminated from wild-type alleles by current CRISPR-Cas systems. Here, we functionally screened six Cas12j nucleas...

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Autores principales: Wang, Yao, Qi, Tao, Liu, Jingtong, Yang, Yuan, Wang, Ziwen, Wang, Ying, Wang, Tianyi, Li, Miaomiao, Li, Mingqing, Lu, Daru, Chang, Alex Chia Yu, Yang, Li, Gao, Song, Wang, Yongming, Lan, Feng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9917002/
https://www.ncbi.nlm.nih.gov/pubmed/36763662
http://dx.doi.org/10.1126/sciadv.abo6405
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author Wang, Yao
Qi, Tao
Liu, Jingtong
Yang, Yuan
Wang, Ziwen
Wang, Ying
Wang, Tianyi
Li, Miaomiao
Li, Mingqing
Lu, Daru
Chang, Alex Chia Yu
Yang, Li
Gao, Song
Wang, Yongming
Lan, Feng
author_facet Wang, Yao
Qi, Tao
Liu, Jingtong
Yang, Yuan
Wang, Ziwen
Wang, Ying
Wang, Tianyi
Li, Miaomiao
Li, Mingqing
Lu, Daru
Chang, Alex Chia Yu
Yang, Li
Gao, Song
Wang, Yongming
Lan, Feng
author_sort Wang, Yao
collection PubMed
description The CRISPR-Cas system can treat autosomal dominant diseases by nonhomologous end joining (NHEJ) gene disruption of mutant alleles. However, many single-nucleotide mutations cannot be discriminated from wild-type alleles by current CRISPR-Cas systems. Here, we functionally screened six Cas12j nucleases and determined Cas12j-8 as an ideal genome editor with a hypercompact size. Cas12j-8 displayed comparable activity to AsCas12a and Un1Cas12f1. Cas12j-8 is a highly specific nuclease sensitive to single-nucleotide mismatches in the protospacer adjacent motif (PAM)–proximal region. We experimentally proved that Cas12j-8 enabled allele-specific disruption of genes with a single-nucleotide polymorphism (SNP). Cas12j-8 recognizes a simple TTN PAM that provides for high target site density. In silico analysis reveals that Cas12j-8 enables allele-specific disruption of 25,931 clinically relevant variants in the ClinVar database, and 485,130,147 SNPs in the dbSNP database. Therefore, Cas12j-8 would be particularly suitable for therapeutic applications.
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spelling pubmed-99170022023-02-11 A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing Wang, Yao Qi, Tao Liu, Jingtong Yang, Yuan Wang, Ziwen Wang, Ying Wang, Tianyi Li, Miaomiao Li, Mingqing Lu, Daru Chang, Alex Chia Yu Yang, Li Gao, Song Wang, Yongming Lan, Feng Sci Adv Biomedicine and Life Sciences The CRISPR-Cas system can treat autosomal dominant diseases by nonhomologous end joining (NHEJ) gene disruption of mutant alleles. However, many single-nucleotide mutations cannot be discriminated from wild-type alleles by current CRISPR-Cas systems. Here, we functionally screened six Cas12j nucleases and determined Cas12j-8 as an ideal genome editor with a hypercompact size. Cas12j-8 displayed comparable activity to AsCas12a and Un1Cas12f1. Cas12j-8 is a highly specific nuclease sensitive to single-nucleotide mismatches in the protospacer adjacent motif (PAM)–proximal region. We experimentally proved that Cas12j-8 enabled allele-specific disruption of genes with a single-nucleotide polymorphism (SNP). Cas12j-8 recognizes a simple TTN PAM that provides for high target site density. In silico analysis reveals that Cas12j-8 enables allele-specific disruption of 25,931 clinically relevant variants in the ClinVar database, and 485,130,147 SNPs in the dbSNP database. Therefore, Cas12j-8 would be particularly suitable for therapeutic applications. American Association for the Advancement of Science 2023-02-10 /pmc/articles/PMC9917002/ /pubmed/36763662 http://dx.doi.org/10.1126/sciadv.abo6405 Text en Copyright © 2023 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). https://creativecommons.org/licenses/by-nc/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (https://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.
spellingShingle Biomedicine and Life Sciences
Wang, Yao
Qi, Tao
Liu, Jingtong
Yang, Yuan
Wang, Ziwen
Wang, Ying
Wang, Tianyi
Li, Miaomiao
Li, Mingqing
Lu, Daru
Chang, Alex Chia Yu
Yang, Li
Gao, Song
Wang, Yongming
Lan, Feng
A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing
title A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing
title_full A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing
title_fullStr A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing
title_full_unstemmed A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing
title_short A highly specific CRISPR-Cas12j nuclease enables allele-specific genome editing
title_sort highly specific crispr-cas12j nuclease enables allele-specific genome editing
topic Biomedicine and Life Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9917002/
https://www.ncbi.nlm.nih.gov/pubmed/36763662
http://dx.doi.org/10.1126/sciadv.abo6405
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