扩大植物基因组编辑范围和配置文件,使用CRISPR-FrCas9系统准Palindromic TA站点
Yao He1,2, Yangshuo Han2, Yanqin Ma1
1Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China, Chengdu, China.
Plant biotechnology journal
|May 7, 2024
概括
来自Faecalibaculum rodentium (FrCas9) 的新CRISPR-Cas9系统提供了增强的植物基因组编辑. 它针对更多的地点,并使精确的基因编辑,大删除和米的基因编辑成为可能,扩大了基因组工程的能力.
科学领域:
- 植物科学 植物科学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 克里斯普尔-Cas9基因组编辑功能强大,但受到原生空间器相邻动机 (PAM) 序列的限制.
- 最常见的CRISPR系统SpCas9需要一个5'-NGG-3' PAM,限制了植物基因组的向.
研究的目的:
- 调查Faecalibaculum rodentium Cas9 (FrCas9) 的植物基因组编辑,特别是在大米中.
- 为了评估FrCas9的效率,PAM的特异性,以及包括大量删除和基础编辑在内的应用.
- 评估FrCas9在植物中的特异性和潜在的非目标效应.
主要方法:
- 在米中的5'-NNTA-3' PAM位点测试了FrCas9的裂变活性.
- TREX2-FrCas9融合是为了增强删除生成而创建的.
- FrCas9被调整为创建细胞基编辑器 (CBEs) 和腺基编辑器 (ABEs).
- 使用全基因组测序分析了非目标突变.
主要成果:
- FrCas9有效地向大米中丰富的5'-NNTA-3' PAM位点,诱导向突变和双基突变.
- 在没有影响编辑效率的情况下,TREX2-FrCas9融合产生了显著更大的删除.
- 从FrCas9衍生出的CBE和ABE成功地引入了针对性的C-to-T和A-to-G基准编辑.
- 全基因组测序表明FrCas9的高特异性,尽管TREX2-FrCas9显示了一些导向RNA独立的目标外突变.
结论:
- CRISPR-FrCas9系统为植物基因组编辑提供了一种高效和多功能工具,包括突变发生,大删除和基因编辑.
- 它对5'-NNTA-3' PAM的识别扩大了植物基因组的准范围.
- FrCas9为先进的植物遗传工程应用提供了一个有前途的替代方案.
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