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Efficient Gene Silencing by Adenine Base Editor-Mediated Start Codon Mutation

Traditional CRISPR/Cas9-based gene knockouts are generated by introducing DNA double-strand breaks (DSBs), but this may cause excessive DNA damage or cell death. CRISPR-based cytosine base editors (CBEs) and adenine base editors (ABEs) can facilitate C-to-T or A-to-G exchanges, respectively, without...

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Autores principales: Wang, Xinjie, Liu, Zhiwei, Li, GuangLei, Dang, Lu, Huang, Shisheng, He, Lei, Ma, Yu’e, Li, Cong, Liu, Ming, Yang, Guang, Huang, Xingxu, Zhou, Fei, Ma, Xiaodong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7001054/
https://www.ncbi.nlm.nih.gov/pubmed/31843453
http://dx.doi.org/10.1016/j.ymthe.2019.11.022
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author Wang, Xinjie
Liu, Zhiwei
Li, GuangLei
Dang, Lu
Huang, Shisheng
He, Lei
Ma, Yu’e
Li, Cong
Liu, Ming
Yang, Guang
Huang, Xingxu
Zhou, Fei
Ma, Xiaodong
author_facet Wang, Xinjie
Liu, Zhiwei
Li, GuangLei
Dang, Lu
Huang, Shisheng
He, Lei
Ma, Yu’e
Li, Cong
Liu, Ming
Yang, Guang
Huang, Xingxu
Zhou, Fei
Ma, Xiaodong
author_sort Wang, Xinjie
collection PubMed
description Traditional CRISPR/Cas9-based gene knockouts are generated by introducing DNA double-strand breaks (DSBs), but this may cause excessive DNA damage or cell death. CRISPR-based cytosine base editors (CBEs) and adenine base editors (ABEs) can facilitate C-to-T or A-to-G exchanges, respectively, without DSBs. CBEs have been employed in a gene knockout strategy: CRISPR-STOP or i-STOP changes single nucleotides to induce in-frame stop codons. However, this strategy is not applicable for some genes, and the unwanted mutations in CBE systems have recently been reported to be surprisingly significant. As a variant, the ABE systems mediate precise editing and have only rare unwanted mutations. In this study, we implemented a new strategy to induce gene silencing (i-Silence) with an ABE-mediated start codon mutation from ATG to GTG or ACG. Using both in vitro and in vivo model systems, we showed that the i-Silence approach is efficient and precise for producing a gene knockout. In addition, the i-Silence strategy can be employed to analyze ~17,804 human genes and can be used to mimic 147 kinds of pathogenic diseases caused by start codon mutations. Altogether, compared to other methods, the ABE-based i-Silence method is a safer gene knockout strategy, and it has promising application potential.
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spelling pubmed-70010542021-02-05 Efficient Gene Silencing by Adenine Base Editor-Mediated Start Codon Mutation Wang, Xinjie Liu, Zhiwei Li, GuangLei Dang, Lu Huang, Shisheng He, Lei Ma, Yu’e Li, Cong Liu, Ming Yang, Guang Huang, Xingxu Zhou, Fei Ma, Xiaodong Mol Ther Original Article Traditional CRISPR/Cas9-based gene knockouts are generated by introducing DNA double-strand breaks (DSBs), but this may cause excessive DNA damage or cell death. CRISPR-based cytosine base editors (CBEs) and adenine base editors (ABEs) can facilitate C-to-T or A-to-G exchanges, respectively, without DSBs. CBEs have been employed in a gene knockout strategy: CRISPR-STOP or i-STOP changes single nucleotides to induce in-frame stop codons. However, this strategy is not applicable for some genes, and the unwanted mutations in CBE systems have recently been reported to be surprisingly significant. As a variant, the ABE systems mediate precise editing and have only rare unwanted mutations. In this study, we implemented a new strategy to induce gene silencing (i-Silence) with an ABE-mediated start codon mutation from ATG to GTG or ACG. Using both in vitro and in vivo model systems, we showed that the i-Silence approach is efficient and precise for producing a gene knockout. In addition, the i-Silence strategy can be employed to analyze ~17,804 human genes and can be used to mimic 147 kinds of pathogenic diseases caused by start codon mutations. Altogether, compared to other methods, the ABE-based i-Silence method is a safer gene knockout strategy, and it has promising application potential. American Society of Gene & Cell Therapy 2020-02-05 2019-11-29 /pmc/articles/PMC7001054/ /pubmed/31843453 http://dx.doi.org/10.1016/j.ymthe.2019.11.022 Text en © 2019 The Author(s) http://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
Wang, Xinjie
Liu, Zhiwei
Li, GuangLei
Dang, Lu
Huang, Shisheng
He, Lei
Ma, Yu’e
Li, Cong
Liu, Ming
Yang, Guang
Huang, Xingxu
Zhou, Fei
Ma, Xiaodong
Efficient Gene Silencing by Adenine Base Editor-Mediated Start Codon Mutation
title Efficient Gene Silencing by Adenine Base Editor-Mediated Start Codon Mutation
title_full Efficient Gene Silencing by Adenine Base Editor-Mediated Start Codon Mutation
title_fullStr Efficient Gene Silencing by Adenine Base Editor-Mediated Start Codon Mutation
title_full_unstemmed Efficient Gene Silencing by Adenine Base Editor-Mediated Start Codon Mutation
title_short Efficient Gene Silencing by Adenine Base Editor-Mediated Start Codon Mutation
title_sort efficient gene silencing by adenine base editor-mediated start codon mutation
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7001054/
https://www.ncbi.nlm.nih.gov/pubmed/31843453
http://dx.doi.org/10.1016/j.ymthe.2019.11.022
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