通过基因表达调节,对Yarrowia lipolytica进行高通量代谢工程
Wei Jiang1, Shengbao Wang1, Daniel Ahlheit1,2
1The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kgs. Lyngby DK-2800, Denmark.
概括
我们开发了TUNEYALI,这是一种CRISPR-Cas9方法,用于*Yarrowia lipolytica*中的高通量基因表达调. 这种方法加速了代谢工程和功能基因组学,因为它可以快速选遗传修饰所需的表型.
科学领域:
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
- 工业微生物学 工业微生物学
背景情况:
- 准确预测所需表型的遗传改变是具有挑战性的.
- 在代谢工程中,需要高通量方法来同时测试多个假设.
研究的目的:
- 介绍TUNEYALI,一种基于CRISPR-Cas9的方法,用于*Yarrowia lipolytica*中的高通量基因表达调.
- 证明TUNEYALI的实用性,用于选影响酵母表型的遗传修饰.
主要方法:
- 开发了一个塑体库,针对56个转录因子 (TF),每个具有7个表达水平.
- 将图书馆转化为Y. lipolytica*菌株,并选出形态,耐热性和贝素生产的变化.
- 利用等离子体测序来识别对观察到的表型变化负责的遗传修饰.
主要成果:
- 确定了增强热耐受性和贝素生产的TF.
- 发现了能够消除伪球体生长的TF.
- 在参考菌株和生产贝坦宁菌株中证明了TUNEYALI的成功应用.
结论:
- TUNE (YALI) 能够高效地对Y. lipolytica*中的基因表达进行高吞吐量调整.
- 这种方法加速了应用菌株开发和功能基因组学研究.
- 可重复使用的库可以为各种基因目标设计,促进更广泛的研究应用.
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