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Updated: Jun 28, 2025

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克里斯普尔-卡斯 (CRISPR-Cas) 工具用于同时对细菌进行转录和翻译控制

Ryan A L Cardiff¹, Ian D Faulkner², Juliana G Beall³

¹Molecular Engineering & Sciences Institute and Center for Synthetic Biology University of Washington Seattle, WA 98195 USA.

Nucleic acids research

|April 13, 2024

概括

CRISPR-dCas13提供了对基因翻译的强有力的控制,克服了细菌操作子中的极性效应. 该系统可为合成生物学和生物技术应用提供精确的基因调节.

科学领域:

微生物合成生物学微生物合成生物学
基因调节基因调节
分子生物学分子生物学

背景情况:

控制特定mRNA点的基因翻译对于合成生物学至关重要.
细菌 (如大肠杆菌) 中的多基因操作子由于极性效应而存在挑战,阻碍了独立的基因调节.
现有的工具很难在没有影响下游基因的情况下精确地准操作子内的单个基因.

研究的目的:

开发和验证CRISPR-dCas13作为微生物系统中精确翻译控制的工具.
为了克服多基因操作子中的极性效应,以实现独立的基因调节.
为了证明转录和翻译控制的联合实用性,以提高基因工程的性能.

主要方法:

使用CRISPR-dCas13系统进行有针对性的翻译抑制.
将dCas13与dCas9.9的极效进行比较.
设计了一种合成的多基因操作子,以使用结合的dCas9转录激活和dCas13转化抑制来证明选择性基因激活.
多重复合的dCas13和dCas9用于增强人乳寡糖的生物合成.

主要成果:

与dCas9.9相比,CRISPR-dCas13的极性效应明显较低 (高达6倍).
通过结合转录和转化控制,在合成操作子中实现单个基因的选择性激活.
成功多重化dCas13和dCas9以改善医学相关化合物的产生.
证明了结合的转录和翻译控制提供了协同的监管效应.

结论:

CRISPR-dCas13为细菌中精确的翻译控制提供了一个强大的工具,减轻极性影响.
转录 (dCas9) 和转化 (dCas13) 控制的结合使复杂的基因调节策略成为可能.
这些先进的工具增强了生物技术中的细菌工程能力,并促进了系统的基因选.

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