SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds

We describe here a CRISPR simultaneous and wide-editing induced by a single system (SWISS), in which RNA aptamers engineered in crRNA scaffold recruit their cognate binding proteins fused with cytidine deaminase and adenosine deaminase to Cas9 nickase target sites to generate multiplexed base editin...

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Autores principales: Li, Chao, Zong, Yuan, Jin, Shuai, Zhu, Haocheng, Lin, Dexing, Li, Shengnan, Qiu, Jin-Long, Wang, Yanpeng, Gao, Caixia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7296638/
https://www.ncbi.nlm.nih.gov/pubmed/32546280
http://dx.doi.org/10.1186/s13059-020-02051-x
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author Li, Chao
Zong, Yuan
Jin, Shuai
Zhu, Haocheng
Lin, Dexing
Li, Shengnan
Qiu, Jin-Long
Wang, Yanpeng
Gao, Caixia
author_facet Li, Chao
Zong, Yuan
Jin, Shuai
Zhu, Haocheng
Lin, Dexing
Li, Shengnan
Qiu, Jin-Long
Wang, Yanpeng
Gao, Caixia
author_sort Li, Chao
collection PubMed
description We describe here a CRISPR simultaneous and wide-editing induced by a single system (SWISS), in which RNA aptamers engineered in crRNA scaffold recruit their cognate binding proteins fused with cytidine deaminase and adenosine deaminase to Cas9 nickase target sites to generate multiplexed base editing. By using paired sgRNAs, SWISS can produce insertions/deletions in addition to base editing. Rice mutants are generated using the SWISS system with efficiencies of cytosine conversion of 25.5%, adenine conversion of 16.4%, indels of 52.7%, and simultaneous triple mutations of 7.3%. The SWISS system provides a powerful tool for multi-functional genome editing in plants.
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spelling pubmed-72966382020-06-16 SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds Li, Chao Zong, Yuan Jin, Shuai Zhu, Haocheng Lin, Dexing Li, Shengnan Qiu, Jin-Long Wang, Yanpeng Gao, Caixia Genome Biol Method We describe here a CRISPR simultaneous and wide-editing induced by a single system (SWISS), in which RNA aptamers engineered in crRNA scaffold recruit their cognate binding proteins fused with cytidine deaminase and adenosine deaminase to Cas9 nickase target sites to generate multiplexed base editing. By using paired sgRNAs, SWISS can produce insertions/deletions in addition to base editing. Rice mutants are generated using the SWISS system with efficiencies of cytosine conversion of 25.5%, adenine conversion of 16.4%, indels of 52.7%, and simultaneous triple mutations of 7.3%. The SWISS system provides a powerful tool for multi-functional genome editing in plants. BioMed Central 2020-06-16 /pmc/articles/PMC7296638/ /pubmed/32546280 http://dx.doi.org/10.1186/s13059-020-02051-x Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Method
Li, Chao
Zong, Yuan
Jin, Shuai
Zhu, Haocheng
Lin, Dexing
Li, Shengnan
Qiu, Jin-Long
Wang, Yanpeng
Gao, Caixia
SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds
title SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds
title_full SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds
title_fullStr SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds
title_full_unstemmed SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds
title_short SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds
title_sort swiss: multiplexed orthogonal genome editing in plants with a cas9 nickase and engineered crispr rna scaffolds
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7296638/
https://www.ncbi.nlm.nih.gov/pubmed/32546280
http://dx.doi.org/10.1186/s13059-020-02051-x
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