CRISPR/Cas-directed programmable assembly of multi-enzyme complexes.,Chemical Communications

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CRISPR/Cas-directed programmable assembly of multi-enzyme complexes.
Chemical Communications ( IF 4.3 ) Pub Date : 2020-05-05 , DOI: 10.1039/d0cc01174f
Samuel Lim ₁ , Jiwoo Kim ₁ , Yujin Kim ₁ , Dawei Xu ₁ , Douglas S Clark ₂

Affiliation

We describe a versatile CRISPR/Cas-based strategy to construct multi-enzyme complexes scaffolded on a DNA template in programmable patterns. Catalytically inactive dCas9 nuclease was used in combination with SpyCatcher-SpyTag chemistry to assemble enzymes in a highly modular fashion. Five enzymes comprising the violacein biosynthesis pathway were precisely organized in nanometer proximity; a notable increase in violacein production demonstrated the benefits of scaffolding.

中文翻译：

CRISPR / Cas指导的多酶复合物的可编程组装。

我们描述了一种通用的基于CRISPR / Cas的策略，以构建可编程模式中DNA模板上支架的多酶复合物。催化不活泼的dCas9核酸酶与SpyCatcher-SpyTag化学结合使用，以高度模块化的方式组装酶。构成紫精素生物合成途径的五种酶在纳米级附近精确组织。紫胶素产量的显着增加证明了脚手架的好处。

更新日期：2020-03-25

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