交换激活模块和双链DNA结合域的合可以促进植物中基于Cas12i3变异的细胞因子和腺因编辑
Chen Zhang1,2, Jingying Li1,2, Yucai Li1,2
1State Key Laboratory of Crop Gene Resources and Breeding, Institute of Crop Sciences (ICS), Chinese Academy of Agricultural Sciences (CAAS), Beijing, 100081, China.
Journal of integrative plant biology
|January 27, 2026
概括
研究人员为植物设计了新的CRISPR/Cas12i3基编辑器,显著提高了米中的C-to-T和A-to-G编辑效率. 这些增强的基础编辑工具为改善作物和开发抗除草剂品种提供了更高的精度.
科学领域:
- 分子生物学分子生物学
- 植物生物技术 植物生物技术
- 基因编辑 基因编辑
背景情况:
- 克里斯普尔/Cas12i3系统提供优势,如较小的蛋白质大小和放松的PAM识别用于基因编辑.
- 现有的基于Cas12i3的细胞氨基基编辑器 (CBEs) 在植物中表现出低效率,而腺氨基基编辑器 (ABEs) 是未经记录的.
- 扩大植物基编辑工具包对于作物改进至关重要.
研究的目的:
- 在大米中设计和评估基于Cas12i3 (5M) 的新型细胞因基编辑器 (CBEs) 和腺因基编辑器 (ABEs).
- 为了提高编辑效率和扩大Cas12i3基础编辑器的编辑窗口.
- 为了证明这些新型编辑器在产生抗除草剂的米的应用.
主要方法:
- 通过将失活的dCas12i3与VP64,Rad51和/或HMG-D域合并,设计基于Cas12i3 (5M) 的CBE和ABE.
- 系统地评估了大米原塑和稳定线的编辑器性能.
- 使用优化编辑器生成了抗除草剂的水生殖质.
主要成果:
- VP64和HMG-D的协同效应组合显著提高了CBE和ABE的效率.
- 基于Cas12i3 (5M) 的优化CBE-V5和ABE-V5在稳定线路中实现了高达4.78倍和3.35倍的编辑效率.
- 最大C-to-T和A-to-G编辑效率达到32.35%和38.24%,在T0代增加了同卵性突变.
结论:
- 基于新型Cas12i3 (5M) 的CBE和ABE系统已成功开发和优化.
- 这些工程编辑器大大丰富了大米和潜在的其他作物的基础编辑工具包.
- 在产生抗除草剂大米方面,已证明实际应用,突出了精密育种的潜力.
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