通过CRISPR/Cas13X辅助可编程和复杂翻译调节,用于控制生物合成
Xianhao Xu1,2, Xueqin Lv1,2, Yanfeng Liu1,2
1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, No. 1800, Lihu Avenue, Binhu District, Wuxi 214122, China.
Nucleic acids research
|January 8, 2025
概括
研究人员开发了基于CRISPR的新型工具,用于在Bacillus subtilis中进行转化基因调节,从而提高微生物细胞工厂的效率. 这些系统在翻译层面上精确控制基因表达,改善有价值化合物的产生.
科学领域:
- 合成生物学 合成生物学
- 分子生物学分子生物学
- 微生物生物技术 微生物生物技术
背景情况:
- 基因调节对于优化微生物细胞工厂至关重要,但目前的工具主要集中在转录控制上.
- 翻译性监管提供了更快的动态响应,但缺乏可编程,高效和多重化的工具.
- 开发新的基因调节策略是推动合成生物学和代谢工程的关键.
研究的目的:
- 开发基于CRISPR的新型系统,用于在Bacillus subtilis的翻译水平上进行可编程的基因调节.
- 使用hfCas13X平台设计CRISPR干扰 (CRISRi) 和CRISPR激活 (CRISRa) 工具.
- 提高这些转化调节系统的效率和适用性,用于代谢工程.
主要方法:
- 使用催化失活的hfCas13X (dhfCas13X) 构建一个CRISPR干扰 (CRISRi) 系统.
- 设计独特的mRNA-crRNA对,以创建降解抑制的CRISPRa (DiCRISPRa) 和翻译启动的CRISPRa (TsCRISPRa) 系统.
- 将dhfCas13X与RNA结合伴侣BHfq融合以提高系统效率.
主要成果:
- 成功开发了CRISPRi和CRISPRa系统,用于在Bacillus subtilis中进行翻译基因调节.
- 在与BHfq.合并时,证明了DiCRISPRa和TsCRISPRa激活效率 (43.2倍) 的显著改善.
- 优化了利博弗拉和2'-糖乳糖生产的代谢网络,分别实现了3倍和1.2倍的标位增加.
结论:
- 开发的CRISPRa和CRISPRi系统为精确的转化基因调节提供了新的工具.
- 这些系统为构建先进的CRISPRa系统和优化微生物细胞工厂提供了新的策略.
- 这些发现推动了基因调节领域及其在生物技术中的应用.
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