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在大米和番茄中高效和可遗传的A-to-K基编辑

Xinbo Li^1,2, Jiyong Xie^3,4, Chao Dong^1,2

¹Ministry of Agriculture and Rural Affairs Key Laboratory of Gene Editing Technologies (Hainan), Institute of Crop Sciences and National Nanfan Research Institute, Chinese Academy of Agricultural Sciences, Sanya, Hainan 572024, China.

Horticulture research

|January 25, 2024

概括

研究人员开发了新的植物基编辑器 (AKBEs),用于高效的A-to-T和A-to-C基因编辑. 这些编辑器扩大了遗传多样性,并使米和西红等作物的基因破坏成为可能.

科学领域:

植物分子生物学植物分子生物学
基因编辑技术的技术
农作物科学农作物科学

背景情况:

细胞因子和腺基编辑器 (CBE和ABE) 是植物基因功能研究和作物改进的关键工具.
现有的基础编辑器可以高效地执行A-to-G和C-to-T/G/A编辑,但在植物中,高效和可遗传的A-to-Y (A-to-T/C) 编辑还不发达.

研究的目的:

构建和评估新的A-to-K基编辑器 (AKBE) 系统,用于单和双植物.
扩大基准编辑器的目标范围,通过整合一个没有PAM的Cas9变体 (nSpRY).
评估A-to-T和A-to-C基础编辑在工厂中的效率和遗传性.

主要方法:

一系列A-to-K基础编辑器 (AKBE) 系统的开发.
整合没有PAM的Cas9变体 (nSpRY) 以扩大目标可访问性.
在米和西红中应用AKBEs,然后对T0和T1代的编辑结果在多个内源位置进行分析.

主要成果:

工厂AKBEs在A-to-G替代中表现出高效率 (平均为41.0%).
实现了显著的A-to-T转换效率 (大米高达25.9%,西红高达10.5%).
优化的AKBE成功产生了A-to-C转换 (平均为1.8%),增强了遗传多样性.
通过准氨酸或氨酸残留物引入了早期停止编码子,显示了基因破坏的潜力.
虽然A-to-T和A-to-C编辑在T0中经常是虚构的,但它们是遗传传给T1后代的.

结论:

开发的AKBE系统对于精确的A-to-T和A-to-C基地编辑在工厂中是有效的.
这些新型编辑器显著扩大了单种和双种植物的基础编辑能力,促进了作物遗传多样性.
AKBEs为基因功能研究和作物育种提供了宝贵的工具,包括有针对性的基因破坏.

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