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基于猪逆转录酶进行哺乳动物基因组编辑的高效主要编辑器

Weiwei Liu¹, Wenxin Duan¹, Zhiwei Peng¹

¹National Key Laboratory for Pig Genetic Improvement and Germplasm Innovation, Ministry of Science and Technology, Jiangxi Agricultural University, Nanchang 330045, China.

Trends in biotechnology

|August 30, 2025

概括

研究人员使用猪反转录酶 (RT) 开发了一种新型主要编辑器 (PE). 这种增强的原始编辑系统 (pvPE) 显示出更高的效率和更少的无意编辑,使猪能够精确编辑基因以进行疾病建模.

科学领域:

分子生物学
遗传学
生物技术

背景情况:

原始编辑是一种强大的基因组编辑技术.
目前的PEs主要使用来自Moloney小鼠白血病病毒 (MMLV) 的逆转录酶 (RT).
需要替代RT源来提高PE的效率和特异性.

研究的目的:

开发一种使用来自猪内源逆转录病毒 (PERV) 的新型主要编辑器 (PE).
优化基于PERV的PE (pvPE) 以提高效率并减少目标外突变.
证明优化的pvPE系统在产生转基因猪的疾病模型中的应用.

主要方法:

来自PERV的RT和结构修改用于增强主编辑器 (PE) 功能.
通过La蛋白的融合来提高PE的活性.
通过代升级 (从pvPE-V1到pvPE-V4) 来优化pvPE系统.
对各种哺乳动物细胞系的pvPE效率和特异性的评估.
评估用于提高pvPE效率的诺科达 (Noc) 治疗.
在猪纤维细胞中同时编辑多个基因.
通过使用pvPE系统生成基因改造的克隆猪.

主要成果:

与pvPE-V1相比,优化的pvPE-V4系统的效率提高了24.38101.69倍.
pvPE-V4的效率高达已有的PE7系统的2.39倍,并且无意编辑的次数显著减少.
诺可达治疗提高了pvPE的平均效率2. 25倍.
在猪纤维细胞中实现了三个基因的同时编辑.
使用pvPE系统成功生成克隆猪,作为人类阿尔茨海默病 (AD) 的潜在模型.

结论:

源自PERV的原始编辑器 (pvPE) 系统代表了基因组编辑技术的重大进步.
与现有系统相比,优化的pvPE,特别是pvPE-V4提供了更高的效率和特异性.
进行多基因编辑和生成疾病模型的能力突显了pvPE在哺乳动物系统中的广泛适用性.

关键词:

阿尔茨海默病在MMLV-RT 其他类型诺科达主要编辑

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