使用CRISPR/Cas9基础编辑器增强Rhodotorula toruloides中的自由脂肪酸生产
1Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.
ACS synthetic biology
|September 1, 2025
概括
研究人员为Rhodotorula toruloides开发了一种基于CRISPR/Cas的基编辑器 (CBE),使得有效的基因编辑成为可能. 这一突破扩大了酵母代谢工程和脂肪酸生产的基因工具.
科学领域:
- 微生物学
- 合成生物学
- 生物技术
背景情况:
- * 罗多托鲁拉多洛伊德是生产脂肪酸等化学物质的有价值的酵母.
- *有限的基因工具限制了它的代谢工程潜力.
- 有效的基因编辑对于释放R. toruloides的能力至关重要.
研究的目的:
- 开发和验证基于CRISPR/Cas的R. toruloides的细胞基编辑器 (CBE) 系统.
- * 为代工程建立多重基因编辑和标记回收.
- * 设计R. toruloides以提高自由脂肪酸的产生.
主要方法:
- * 在R. toruloides中开发基于CRISPR/Cas的基编辑器 (CBE).
- * 集成可诱导的Cre-loxP系统用于选择标记回收.
- * CBE用于单基因和多基因干扰,包括参与脂质代谢的基因.
主要成果:
- 使用CBE系统实现了高达90%的单基因破坏效率.
- * 证明成功地同时破坏了四个基因,效率为5%.
- * 实现了选择标记的回收,效率超过70%,并构建了 uracil-auxotrophic 菌株.
- 通过破坏脂质代谢基因产生512.3毫克/升的菌株.
结论:
- * 开发的CBE系统是R. toruloides的一个精确和特定的基因组编辑工具.
- * 这一工具集显著扩大了R. toruloides的基因工程能力.
- 这项研究为增强R. toruloides的代谢工程和化学生产铺平了道路.
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