Highly efficient prime editors for mammalian genome editing based on porcine retrovirus reverse transcriptase.

IF 14.9 1区工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Trends in biotechnology Pub Date : 2025-08-30 DOI:10.1016/j.tibtech.2025.07.029

Weiwei Liu, Wenxin Duan, Zhiwei Peng, Yaya Liao, Xiaoguo Wang, Ruirong Liu, Qiqi Jing, Haoyun Jiang, Yuhang Fan, Liming Ge, Lusheng Huang, Yuyun Xing

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引用次数: 0

Abstract

Prime editing is a versatile and precise genome-editing tool. Most prime editors (PEs) rely on reverse transcriptase (RT) derived from Moloney murine leukemia virus (MMLV). Here, we established a PE, pvPE, using a RT derived from a porcine endogenous retrovirus (PERV) from a Bama mini-pig. Through various optimization strategies, including RT engineering, structural modifications, and La protein fusion, we gradually upgraded to pvPE-V4. This version achieved 24.38-101.69-fold higher efficiency compared with pvPE-V1 and up to 2.39-fold higher efficiency than another upgraded PE, PE7, with significantly fewer unintended edits across multiple mammalian cell lines. We further show that nocodazole (Noc) significantly enhanced pvPE efficiency by 2.25-fold on average. Using our pvPE system, we efficiently modified three genes simultaneously in porcine fibroblasts and subsequently generated cloned pigs that could serve as valuable models for Alzheimer's disease (AD) in humans. Our results highlight the broad application prospects of pvPE systems in mammalian genome editing.

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基于猪逆转录病毒逆转录酶的高效哺乳动物基因组编辑器。

启动编辑是一种多功能和精确的基因组编辑工具。大多数启动编辑器（PEs）依赖于来自Moloney小鼠白血病病毒（MMLV）的逆转录酶（RT）。在这里，我们使用来自巴马迷你猪的猪内源性逆转录病毒（PERV）衍生的RT建立了PE， pvPE。通过RT工程、结构修改、La蛋白融合等多种优化策略，逐步升级为pvPE-V4。与ppe - v1相比，该版本的效率提高了24.38-101.69倍，比另一种升级PE PE7的效率提高了2.39倍，并且在多个哺乳动物细胞系中显著减少了意外编辑。我们进一步表明，nocodazole （Noc）显著提高了pvPE效率，平均提高了2.25倍。利用我们的pvPE系统，我们有效地同时修饰了猪成纤维细胞中的三个基因，随后产生了克隆猪，可以作为人类阿尔茨海默病（AD）的有价值的模型。我们的研究结果突出了pvPE系统在哺乳动物基因组编辑中的广阔应用前景。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Trends in biotechnology 工程技术-生物工程与应用微生物

CiteScore

28.60

自引率

1.20%

发文量

198

审稿时长

1 months

期刊介绍： Trends in Biotechnology publishes reviews and perspectives on the applied biological sciences, focusing on useful science applied to, derived from, or inspired by living systems. The major themes that TIBTECH is interested in include: Bioprocessing (biochemical engineering, applied enzymology, industrial biotechnology, biofuels, metabolic engineering) Omics (genome editing, single-cell technologies, bioinformatics, synthetic biology) Materials and devices (bionanotechnology, biomaterials, diagnostics/imaging/detection, soft robotics, biosensors/bioelectronics) Therapeutics (biofabrication, stem cells, tissue engineering and regenerative medicine, antibodies and other protein drugs, drug delivery) Agroenvironment (environmental engineering, bioremediation, genetically modified crops, sustainable development).