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协同优化增强了树中细胞氨基基编辑器的精度和效率

Han Liu^1,2,3,4, Mengyu Zhang^1,2,3,4, Leiqian Sun^1,2,3,4

¹State Key Laboratory of Tree Genetics and Breeding, Beijing Forestry University, Beijing, China.

Communications biology

|June 10, 2025

概括

我们开发了hyPopCBE-V4,这是一款高效的树细胞酶基编辑器 (CBE),显著提高了C-to-T编辑的准确性和效率. 这一进步使得像树和其他木质植物的除草剂耐药性等特征的精确基因修饰成为可能.

科学领域:

植物生物技术植物生物技术
基因组编辑基因组编辑
分子生物学分子生物学

背景情况:

克里斯普尔/卡斯9细胞酶基编辑 (CBE) 提供了精确的植物基因组修改.
现有的CBE系统显示低效率和不精确的编辑.
为树优化CBE对于农业和林业应用至关重要.

研究的目的:

设计一个高效率的树CBE系统 (hyPopCBE).
优化CBE系统以提高树的效率和精度.
使用增强的CBE系统开发抗除草剂的子生殖质.

主要方法:

集成了MS2-UGI系统,Rad51DNA结合域,并修改了核定位信号.
开发了逐步优化,导致hyPopCBE-V4系统.
诱导了树PagalS基因中的Pro197Leu突变.

主要成果:

hyPopCBE-V4在木质植物中表现出协同效应,提高了C-to-T编辑效率.
干净的C-to-T编辑从20.93%增加到40.48%,干净的同卵性编辑效率从4.65%增加到21.43%.
经过编辑的PagALS基因的工程木线显示出对tribenuron和nicosulfuron的高耐药性.

结论:

多组件协同优化策略显著提高了CBE的效率和精度.
hyPopCBE-V4为和其他林业物种的基因改造提供了更精确,更有效的工具.
成功生成了抗除草剂的子生殖质,证明了该系统在育种计划中的实际应用.

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