CRISPR/Cas-directed programmable assembly of multi-enzyme complexes
Samuel Lim, Jiwoo Kim, Yu‐Jin Kim, Dawei Xu, Douglas S. Clark
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.
Topics & Concepts
CRISPRConstruct (python library)Combinatorial chemistryEnzymeComputer scienceChemistryComputational biologyNanotechnologyBiochemistryBiologyMaterials scienceProgramming languageGeneCRISPR and Genetic EngineeringAdvanced biosensing and bioanalysis techniquesRNA and protein synthesis mechanisms