PE6c greatly enhances prime editing in transgenic rice plants

IF 9.3 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Zhenghong Cao, Wei Sun, Dexin Qiao, Junya Wang, Siyun Li, Xiaohan Liu, Cuiping Xin, Yu Lu, Syeda Leeda Gul, Xue-Chen Wang, Qi-Jun Chen
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Abstract

Prime editing is a versatile CRISPR/Cas-based precise genome-editing technique for crop breeding. Four new types of prime editors (PEs) named PE6a–d were recently generated using evolved and engineered reverse transcriptase (RT) variants from three different sources. In this study, we tested the editing efficiencies of four PE6 variants and two additional PE6 constructs with double-RT modules in transgenic rice (Oryza sativa) plants. PE6c, with an evolved and engineered RT variant from the yeast Tf1 retrotransposon, yielded the highest prime-editing efficiency. The average fold change in the editing efficiency of PE6c compared with PEmax exceeded 3.5 across 18 agronomically important target sites from 15 genes. We also demonstrated the feasibility of using two RT modules to improve prime-editing efficiency. Our results suggest that PE6c or its derivatives would be an excellent choice for prime editing in monocot plants. In addition, our findings have laid a foundation for prime-editing-based breeding of rice varieties with enhanced agronomically important traits.

Abstract Image

PE6c 可大大增强转基因水稻植株的素体编辑能力。
主编辑是一种基于 CRISPR/Cas 的多功能精准基因组编辑技术,可用于作物育种。最近,利用来自三个不同来源的进化和工程化反转录酶(RT)变体生成了四种新型的主编辑器(PE),分别命名为 PE6a-d。在本研究中,我们在转基因水稻(Oryza sativa)植株中测试了四种 PE6 变体和另外两种带有双 RT 模块的 PE6 构建体的编辑效率。含有从酵母 Tf1 反转座子进化和工程化的 RT 变体的 PE6c 产生了最高的质粒编辑效率。与 PEmax 相比,PE6c 在 15 个基因的 18 个重要农艺目标位点上的编辑效率平均折叠变化超过 3.5。我们还证明了使用两个 RT 模块提高质粒编辑效率的可行性。我们的研究结果表明,PE6c 或其衍生物将是单子叶植物质粒编辑的绝佳选择。此外,我们的研究结果还为基于素编辑技术培育具有更多重要农艺性状的水稻品种奠定了基础。
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来源期刊
Journal of Integrative Plant Biology
Journal of Integrative Plant Biology 生物-生化与分子生物学
CiteScore
18.00
自引率
5.30%
发文量
220
审稿时长
3 months
期刊介绍: Journal of Integrative Plant Biology is a leading academic journal reporting on the latest discoveries in plant biology.Enjoy the latest news and developments in the field, understand new and improved methods and research tools, and explore basic biological questions through reproducible experimental design, using genetic, biochemical, cell and molecular biological methods, and statistical analyses.
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