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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...

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Detalles Bibliográficos
Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2023
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.
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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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