复子介导的多重基因组编辑和埃舍里奇亚大肠杆菌的持续进化
Wenqian Liu1,2, Siqi Zuo1,2, Youran Shao1,2
1Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
Nucleic acids research
|July 20, 2023
概括
我们开发了一种新型的逆子介导基因组编辑系统 (REGES),用于高效的多重基因组编辑. 这种多功能工具可以实现动态和同时的突变发生,用于合成生物学和代谢工程中的应用.
科学领域:
- 合成生物学 合成生物学
- 分子生物学分子生物学
- 基因组工程是基因组工程.
背景情况:
- 现有的基因组编辑方法对 prokaryotes 中的动态和同时突变发生有局限性.
- 需要多功能工具来高效地修改多个基因组位点.
研究的目的:
- 开发和优化一种新的多重基因组编辑系统,用于 prokaryotes.
- 为了证明系统在生成变体库和连续蛋白质进化的实用性.
主要方法:
- 开发和优化逆子介导基因组编辑系统 (REGES).
- 应用REGES用于单,双,三,四个位置的基因组编辑.
- 使用退化的RBS序列生成聚合和条码变体库.
- 通过结合反子,基编辑和易出错的转录来证明持续的体内蛋白质进化.
主要成果:
- 实现了高的编辑效率:~100% (单),85% (双),69% (三),和25% (四) 的位置编辑.
- 成功生成了变体库,以微调基因表达,以改善乙醇耐受性和生物生物合成.
- 对于持续的体内蛋白质进化的证明 REGES.
结论:
- REGES 是一种强大而通用的工具,用于多重基因组编辑.
- 该系统促进了对合成生物学和代谢工程应用的变体库的生成和持续演变.
- 在微生物系统中推进基因工程方面,REGES具有广泛的适用性.
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