CRISPR‐BETS: a base‐editing design tool for generating stop codons

Cytosine base editors (CBEs) can install a predefined stop codon at the target site, representing a more predictable and neater method for creating genetic knockouts without altering the genome size. Due to the enhanced predictability of the editing outcomes, it is also more efficient to obtain homo...

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Autores principales: Wu, Yuechao, He, Yao, Sretenovic, Simon, Liu, Shishi, Cheng, Yanhao, Han, Yangshuo, Liu, Guanqing, Bao, Yu, Fang, Qing, Zheng, Xuelian, Zhou, Jianping, Qi, Yiping, Zhang, Yong, Zhang, Tao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8882796/
https://www.ncbi.nlm.nih.gov/pubmed/34669232
http://dx.doi.org/10.1111/pbi.13732
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author Wu, Yuechao
He, Yao
Sretenovic, Simon
Liu, Shishi
Cheng, Yanhao
Han, Yangshuo
Liu, Guanqing
Bao, Yu
Fang, Qing
Zheng, Xuelian
Zhou, Jianping
Qi, Yiping
Zhang, Yong
Zhang, Tao
author_facet Wu, Yuechao
He, Yao
Sretenovic, Simon
Liu, Shishi
Cheng, Yanhao
Han, Yangshuo
Liu, Guanqing
Bao, Yu
Fang, Qing
Zheng, Xuelian
Zhou, Jianping
Qi, Yiping
Zhang, Yong
Zhang, Tao
author_sort Wu, Yuechao
collection PubMed
description Cytosine base editors (CBEs) can install a predefined stop codon at the target site, representing a more predictable and neater method for creating genetic knockouts without altering the genome size. Due to the enhanced predictability of the editing outcomes, it is also more efficient to obtain homozygous mutants in the first generation. With the recent advancement of CBEs on improved editing activity, purify and specificity in plants and animals, base editing has become a more appealing technology for generating knockouts. However, there is a lack of design tools that can aid the adoption of CBEs for achieving such a purpose, especially in plants. Here, we developed a user‐friendly design tool named CRISPR‐BETS (base editing to stop), which helps with guide RNA (gRNA) design for introducing stop codons in the protein‐coding genes of interest. We demonstrated in rice and tomato that CRISPR‐BETS is easy‐to‐use, and its generated gRNAs are highly specific and efficient for generating stop codons and obtaining homozygous knockout lines. While we tailored the tool for the plant research community, CRISPR‐BETS can also serve non‐plant species.
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spelling pubmed-88827962022-03-04 CRISPR‐BETS: a base‐editing design tool for generating stop codons Wu, Yuechao He, Yao Sretenovic, Simon Liu, Shishi Cheng, Yanhao Han, Yangshuo Liu, Guanqing Bao, Yu Fang, Qing Zheng, Xuelian Zhou, Jianping Qi, Yiping Zhang, Yong Zhang, Tao Plant Biotechnol J Research Articles Cytosine base editors (CBEs) can install a predefined stop codon at the target site, representing a more predictable and neater method for creating genetic knockouts without altering the genome size. Due to the enhanced predictability of the editing outcomes, it is also more efficient to obtain homozygous mutants in the first generation. With the recent advancement of CBEs on improved editing activity, purify and specificity in plants and animals, base editing has become a more appealing technology for generating knockouts. However, there is a lack of design tools that can aid the adoption of CBEs for achieving such a purpose, especially in plants. Here, we developed a user‐friendly design tool named CRISPR‐BETS (base editing to stop), which helps with guide RNA (gRNA) design for introducing stop codons in the protein‐coding genes of interest. We demonstrated in rice and tomato that CRISPR‐BETS is easy‐to‐use, and its generated gRNAs are highly specific and efficient for generating stop codons and obtaining homozygous knockout lines. While we tailored the tool for the plant research community, CRISPR‐BETS can also serve non‐plant species. John Wiley and Sons Inc. 2021-11-02 2022-03 /pmc/articles/PMC8882796/ /pubmed/34669232 http://dx.doi.org/10.1111/pbi.13732 Text en © 2021 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Wu, Yuechao
He, Yao
Sretenovic, Simon
Liu, Shishi
Cheng, Yanhao
Han, Yangshuo
Liu, Guanqing
Bao, Yu
Fang, Qing
Zheng, Xuelian
Zhou, Jianping
Qi, Yiping
Zhang, Yong
Zhang, Tao
CRISPR‐BETS: a base‐editing design tool for generating stop codons
title CRISPR‐BETS: a base‐editing design tool for generating stop codons
title_full CRISPR‐BETS: a base‐editing design tool for generating stop codons
title_fullStr CRISPR‐BETS: a base‐editing design tool for generating stop codons
title_full_unstemmed CRISPR‐BETS: a base‐editing design tool for generating stop codons
title_short CRISPR‐BETS: a base‐editing design tool for generating stop codons
title_sort crispr‐bets: a base‐editing design tool for generating stop codons
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8882796/
https://www.ncbi.nlm.nih.gov/pubmed/34669232
http://dx.doi.org/10.1111/pbi.13732
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