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Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase
Prime editor (PE) is a versatile genome editing tool that does not need extra DNA donors or inducing double-strand breaks. However, in vivo implementation of PE remains a challenge because of its oversized composition. In this study, we screened out the smallest truncated Moloney murine leukemia vir...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10480570/ https://www.ncbi.nlm.nih.gov/pubmed/37680986 http://dx.doi.org/10.1016/j.omtn.2023.08.018 |
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author | Lan, Ting Chen, Huangyao Tang, Chengcheng Wei, Yuhui Liu, Yang Zhou, Jizeng Zhuang, Zhenpeng Zhang, Quanjun Chen, Min Zhou, Xiaoqing Chi, Yue Wang, Jinling He, Yu Lai, Liangxue Zou, Qingjian |
author_facet | Lan, Ting Chen, Huangyao Tang, Chengcheng Wei, Yuhui Liu, Yang Zhou, Jizeng Zhuang, Zhenpeng Zhang, Quanjun Chen, Min Zhou, Xiaoqing Chi, Yue Wang, Jinling He, Yu Lai, Liangxue Zou, Qingjian |
author_sort | Lan, Ting |
collection | PubMed |
description | Prime editor (PE) is a versatile genome editing tool that does not need extra DNA donors or inducing double-strand breaks. However, in vivo implementation of PE remains a challenge because of its oversized composition. In this study, we screened out the smallest truncated Moloney murine leukemia virus (MMLV) reverse transcriptase (RT) with the F155Y mutation to keep gene editing efficiency. We discovered the most efficient gene editing variants of MMLV RT with the smallest size. After optimization of the pegRNAs and incorporation with nick sgRNAs, the mini-PE delivered up to 10% precise editing at target sites in human and mouse cells. It also edited the mouse Hsf1 gene in the mouse retina precisely after delivery with adeno-associated viruses (AAVs), although the editing efficiency was lower than 1%. We will focus on improving the editing efficiency of mini-PE and exploiting its therapeutic potential against human genetic diseases. |
format | Online Article Text |
id | pubmed-10480570 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-104805702023-09-07 Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase Lan, Ting Chen, Huangyao Tang, Chengcheng Wei, Yuhui Liu, Yang Zhou, Jizeng Zhuang, Zhenpeng Zhang, Quanjun Chen, Min Zhou, Xiaoqing Chi, Yue Wang, Jinling He, Yu Lai, Liangxue Zou, Qingjian Mol Ther Nucleic Acids Brief Report Prime editor (PE) is a versatile genome editing tool that does not need extra DNA donors or inducing double-strand breaks. However, in vivo implementation of PE remains a challenge because of its oversized composition. In this study, we screened out the smallest truncated Moloney murine leukemia virus (MMLV) reverse transcriptase (RT) with the F155Y mutation to keep gene editing efficiency. We discovered the most efficient gene editing variants of MMLV RT with the smallest size. After optimization of the pegRNAs and incorporation with nick sgRNAs, the mini-PE delivered up to 10% precise editing at target sites in human and mouse cells. It also edited the mouse Hsf1 gene in the mouse retina precisely after delivery with adeno-associated viruses (AAVs), although the editing efficiency was lower than 1%. We will focus on improving the editing efficiency of mini-PE and exploiting its therapeutic potential against human genetic diseases. American Society of Gene & Cell Therapy 2023-08-18 /pmc/articles/PMC10480570/ /pubmed/37680986 http://dx.doi.org/10.1016/j.omtn.2023.08.018 Text en © 2023 The Author(s) https://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 | Brief Report Lan, Ting Chen, Huangyao Tang, Chengcheng Wei, Yuhui Liu, Yang Zhou, Jizeng Zhuang, Zhenpeng Zhang, Quanjun Chen, Min Zhou, Xiaoqing Chi, Yue Wang, Jinling He, Yu Lai, Liangxue Zou, Qingjian Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase |
title | Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase |
title_full | Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase |
title_fullStr | Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase |
title_full_unstemmed | Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase |
title_short | Mini-PE, a prime editor with compact Cas9 and truncated reverse transcriptase |
title_sort | mini-pe, a prime editor with compact cas9 and truncated reverse transcriptase |
topic | Brief Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10480570/ https://www.ncbi.nlm.nih.gov/pubmed/37680986 http://dx.doi.org/10.1016/j.omtn.2023.08.018 |
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