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WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing

Large scale genomic aberrations including duplication, deletion, translocation, and other structural changes are the cause of a subtype of hereditary genetic disorders and contribute to onset or progress of cancer. The current prime editor, PE2, consisting of Cas9-nickase and reverse transcriptase e...

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Autores principales: Tao, Rui, Wang, Yanhong, Hu, Yun, Jiao, Yaoge, Zhou, Lifang, Jiang, Lurong, Li, Li, He, Xingyu, Li, Min, Yu, Yamei, Chen, Qiang, Yao, Shaohua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9018734/
https://www.ncbi.nlm.nih.gov/pubmed/35440051
http://dx.doi.org/10.1038/s41392-022-00936-w
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author Tao, Rui
Wang, Yanhong
Hu, Yun
Jiao, Yaoge
Zhou, Lifang
Jiang, Lurong
Li, Li
He, Xingyu
Li, Min
Yu, Yamei
Chen, Qiang
Yao, Shaohua
author_facet Tao, Rui
Wang, Yanhong
Hu, Yun
Jiao, Yaoge
Zhou, Lifang
Jiang, Lurong
Li, Li
He, Xingyu
Li, Min
Yu, Yamei
Chen, Qiang
Yao, Shaohua
author_sort Tao, Rui
collection PubMed
description Large scale genomic aberrations including duplication, deletion, translocation, and other structural changes are the cause of a subtype of hereditary genetic disorders and contribute to onset or progress of cancer. The current prime editor, PE2, consisting of Cas9-nickase and reverse transcriptase enables efficient editing of genomic deletion and insertion, however, at small scale. Here, we designed a novel prime editor by fusing reverse transcriptase (RT) to nuclease wild-type Cas9 (WT-PE) to edit large genomic fragment. WT-PE system simultaneously introduced a double strand break (DSB) and a single 3′ extended flap in the target site. Coupled with paired prime editing guide RNAs (pegRNAs) that have complementary sequences in their 3′ terminus while target different genomic regions, WT-PE produced bi-directional prime editing, which enabled efficient and versatile large-scale genome editing, including large fragment deletion up to 16.8 megabase (Mb) pairs and chromosomal translocation. Therefore, our WT-PE system has great potential to model or treat diseases related to large-fragment aberrations.
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spelling pubmed-90187342022-04-28 WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing Tao, Rui Wang, Yanhong Hu, Yun Jiao, Yaoge Zhou, Lifang Jiang, Lurong Li, Li He, Xingyu Li, Min Yu, Yamei Chen, Qiang Yao, Shaohua Signal Transduct Target Ther Article Large scale genomic aberrations including duplication, deletion, translocation, and other structural changes are the cause of a subtype of hereditary genetic disorders and contribute to onset or progress of cancer. The current prime editor, PE2, consisting of Cas9-nickase and reverse transcriptase enables efficient editing of genomic deletion and insertion, however, at small scale. Here, we designed a novel prime editor by fusing reverse transcriptase (RT) to nuclease wild-type Cas9 (WT-PE) to edit large genomic fragment. WT-PE system simultaneously introduced a double strand break (DSB) and a single 3′ extended flap in the target site. Coupled with paired prime editing guide RNAs (pegRNAs) that have complementary sequences in their 3′ terminus while target different genomic regions, WT-PE produced bi-directional prime editing, which enabled efficient and versatile large-scale genome editing, including large fragment deletion up to 16.8 megabase (Mb) pairs and chromosomal translocation. Therefore, our WT-PE system has great potential to model or treat diseases related to large-fragment aberrations. Nature Publishing Group UK 2022-04-20 /pmc/articles/PMC9018734/ /pubmed/35440051 http://dx.doi.org/10.1038/s41392-022-00936-w Text en © The Author(s) 2022 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Tao, Rui
Wang, Yanhong
Hu, Yun
Jiao, Yaoge
Zhou, Lifang
Jiang, Lurong
Li, Li
He, Xingyu
Li, Min
Yu, Yamei
Chen, Qiang
Yao, Shaohua
WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing
title WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing
title_full WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing
title_fullStr WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing
title_full_unstemmed WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing
title_short WT-PE: Prime editing with nuclease wild-type Cas9 enables versatile large-scale genome editing
title_sort wt-pe: prime editing with nuclease wild-type cas9 enables versatile large-scale genome editing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9018734/
https://www.ncbi.nlm.nih.gov/pubmed/35440051
http://dx.doi.org/10.1038/s41392-022-00936-w
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