在Acremonium chrysogenum中使用高效的5S rRNA-CRISPR/Cas9基因组编辑工具
Xiaomei Zheng1,2,3,4,5, Yanjiao Zhai1,2, Hetti Arachchige Wathsalie Chathurika1,2
1College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China.
Journal of agricultural and food chemistry
|September 2, 2025
概括
一个新的CRISPR/Cas9系统用于*Acremonium chrysogenum*使用内源5SrRNA促进剂进行简化和高效的基因组编辑. 这种强大的工具能够精确地删除基因和大量染色体,
科学领域:
- 微生物学
- 分子生物学
- 生物技术
背景情况:
- 阿克雷莫尼亚是一种主要的工业化素C (CPC) 生产者.
- 在A. chrysogenum中现有的CRISPR/Cas9系统依赖于异质促进体,需要额外的sgRNA处理元素,增加复杂性.
- 开发高效的基因组编辑工具对于A. chrysogenum的代谢工程和菌株改进至关重要.
研究的目的:
- 开发一种简化且高效的CRISPR/Cas9基因组编辑系统.
- 使用内源5SrRNA促进体进行sgRNA转录,从而消除了处理元素的需要.
- 证明该系统在基因破坏,单基因删除,大规模染色体删除和代基因编辑方面的有效性.
主要方法:
- 在A. chrysogenum中开发使用内源5SrRNA促进体的CRISPR/Cas9系统.
- 针对SORB基因进行基因破坏.
- 在sorbicillinoid生物合成基因群中执行单基因删除和大规模染色体删除 (高达66.17kb).
- 在kusA位点实施同源重组介导标记替代以实现精确和代的基因编辑.
主要成果:
- 达到100%的基因破坏效率, 针对SORB基因.
- 证明了100%的单基因删除效率.
- 在没有捐赠DNA的情况下成功进行了大规模的染色体删除.
- 通过同源重组促进精确和代基因编辑.
结论:
- 开发的5S rRNA-CRISPR/Cas9系统是一种简化,强大和高效的基因组编辑工具包.
- 这种系统避免了需要额外的sgRNA处理元素,减少了实验的复杂性.
- 它可以实现多功能应用,包括功能基因组学,菌株改进和A. chrysogenum的大规模基因组工程.
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