使用益生菌来源的CRISPR-Cas9系统进行高效的植物基因组工程
Zhaohui Zhong1,2, Guanqing Liu3,4,5, Zhongjie Tang1
1Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China, 610054, Chengdu, China.
Nature communications
|September 29, 2023
概括
研究人员开发了一种新的CRISPR-Cas9基因组编辑系统,从植物中使用Lactobacillus rhamnosus (LrCas9). 这种高效的工具超越了现有的系统在编辑各种作物,如大米和小麦.
科学领域:
- 分子生物学分子生物学
- 植物科学 植物科学
- 生物技术是生物技术.
背景情况:
- 链球菌 pyogenes Cas9 (SpCas9) 是最常见的CRISPR-Cas基因组编辑系统.
- SpCas9源自人类的病原体,促使人们寻找替代系统.
- 在植物生物技术中需要高效和多功能基因组工程工具.
研究的目的:
- 建立一个高效的植物基因组工程系统,使用来自Lactobacillus rhamnosus (LrCas9) 的CRISPR-Cas9.
- 为了验证LrCas9.9的5'-NGAAA-3'原位器相邻动机 (PAM).
- 为了证明LrCas9在各种植物物种和应用中的编辑效率和特异性.
主要方法:
- 在形数据挖掘以确定潜在的CRISPR-Cas9系统.
- 细菌PAM消耗测定以确认LrCas9 PAM序列.
- 在大米,小麦,西红和Larix细胞中进行基因组编辑实验.
- 开发LrCas9衍生基编辑器和CRISPR干扰/激活系统.
主要成果:
- 在大米,小麦,西红和Larix细胞中,LrCas9表现出卓越的编辑效率,其性能优于LbCas12a,SpCas9-NG和SpRY.
- 多复合基因淘汰和基因淘汰在稳定的大米系中使用LrCas9.9实现.
- 通过有针对性的促进体删除和编码序列编辑,LrCas9的高特异性得到证明.
- 成功开发了由LrCas9衍生的细胞因子和腺因基编辑器.
- 有效的CRISPR干扰和激活系统是通过利用LrCas9的A/T丰富的PAM准来创建的.
结论:
- CRISPR-LrCas9是一种高效且易于使用的植物基因组工程工具.
- 与现有系统相比,LrCas9提供了优势,包括更广泛的PAM定位和更高的效率.
- 该系统扩大了作物中的基因组编辑能力,并有可能用于各种应用.
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