Efficient and versatile multiplex prime editing in hexaploid wheat

Prime editing is limited by low efficiency in plants. Here, we develop an upgraded engineered plant prime editor in hexaploid wheat, ePPEplus, by introducing a V223A substitution into reverse transcriptase in the ePPEmax* architecture. ePPEplus enhances the efficiency by an average 33.0-fold and 6.4...

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Autores principales: Ni, Pei, Zhao, Yidi, Zhou, Ximeng, Liu, Zehua, Huang, Zhengwei, Ni, Zhongfu, Sun, Qixin, Zong, Yuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Method
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10308706/
https://www.ncbi.nlm.nih.gov/pubmed/37386475
http://dx.doi.org/10.1186/s13059-023-02990-1
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author Ni, Pei
Zhao, Yidi
Zhou, Ximeng
Liu, Zehua
Huang, Zhengwei
Ni, Zhongfu
Sun, Qixin
Zong, Yuan
author_facet Ni, Pei
Zhao, Yidi
Zhou, Ximeng
Liu, Zehua
Huang, Zhengwei
Ni, Zhongfu
Sun, Qixin
Zong, Yuan
author_sort Ni, Pei
collection PubMed
description Prime editing is limited by low efficiency in plants. Here, we develop an upgraded engineered plant prime editor in hexaploid wheat, ePPEplus, by introducing a V223A substitution into reverse transcriptase in the ePPEmax* architecture. ePPEplus enhances the efficiency by an average 33.0-fold and 6.4-fold compared to the original PPE and ePPE, respectively. Importantly, a robust multiplex prime editing platform is established for simultaneous editing of four to ten genes in protoplasts and up to eight genes in regenerated wheat plants at frequencies up to 74.5%, thus expanding the applicability of prime editors for stacking of multiple agronomic traits. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-023-02990-1.
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spelling pubmed-103087062023-06-30 Efficient and versatile multiplex prime editing in hexaploid wheat Ni, Pei Zhao, Yidi Zhou, Ximeng Liu, Zehua Huang, Zhengwei Ni, Zhongfu Sun, Qixin Zong, Yuan Genome Biol Method Prime editing is limited by low efficiency in plants. Here, we develop an upgraded engineered plant prime editor in hexaploid wheat, ePPEplus, by introducing a V223A substitution into reverse transcriptase in the ePPEmax* architecture. ePPEplus enhances the efficiency by an average 33.0-fold and 6.4-fold compared to the original PPE and ePPE, respectively. Importantly, a robust multiplex prime editing platform is established for simultaneous editing of four to ten genes in protoplasts and up to eight genes in regenerated wheat plants at frequencies up to 74.5%, thus expanding the applicability of prime editors for stacking of multiple agronomic traits. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13059-023-02990-1. BioMed Central 2023-06-29 /pmc/articles/PMC10308706/ /pubmed/37386475 http://dx.doi.org/10.1186/s13059-023-02990-1 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This 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/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://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
Ni, Pei
Zhao, Yidi
Zhou, Ximeng
Liu, Zehua
Huang, Zhengwei
Ni, Zhongfu
Sun, Qixin
Zong, Yuan
Efficient and versatile multiplex prime editing in hexaploid wheat
title Efficient and versatile multiplex prime editing in hexaploid wheat
title_full Efficient and versatile multiplex prime editing in hexaploid wheat
title_fullStr Efficient and versatile multiplex prime editing in hexaploid wheat
title_full_unstemmed Efficient and versatile multiplex prime editing in hexaploid wheat
title_short Efficient and versatile multiplex prime editing in hexaploid wheat
title_sort efficient and versatile multiplex prime editing in hexaploid wheat
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10308706/
https://www.ncbi.nlm.nih.gov/pubmed/37386475
http://dx.doi.org/10.1186/s13059-023-02990-1
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