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CRISPR/Cas-directed programmable assembly of multi-enzyme complexes

Abstract

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.

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Supplementary files

Article information


Submitted
14 Feb 2020
Accepted
25 Mar 2020
First published
25 Mar 2020

Chem. Commun., 2020, Accepted Manuscript
Article type
Communication

CRISPR/Cas-directed programmable assembly of multi-enzyme complexes

S. Lim, J. Kim, Y. Kim, D. Xu and D. S. Clark, Chem. Commun., 2020, Accepted Manuscript , DOI: 10.1039/D0CC01174F

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