Team Competition
iGEM 2021
International Genetically Engineered Machine Competition
Overview
Background
Vibrio natriegens is a promising chassis for synthetic biology because of its exceptionally rapid doubling time of less than 10 minutes, which is approximately twice as fast as that of Escherichia coli, and its high ribosome content per cell. Yet despite these favourable attributes, the organism remains comparatively underdeveloped as an engineering platform. Regulatory parts are limited, CRISPR based tools are largely unavailable, and V. natriegens appears particularly susceptible to the burden associated with heterologous gene expression. Our project aims to help close this gap by establishing a comprehensive and tunable toolkit for gene regulation in V. natriegens.
What We Did
- SSR Part Collection: Built a Golden Gate-compatible library of key regulatory elements, including promoters, RBSs, insulators, ORIs, and inducible systems, to enable fine-grained gene expression control in V. natriegens.
- Lightweight CRISPR Toolkit: Developed CRISPRa and CRISPRi systems using compact Cas proteins to minimize metabolic burden, optimizing both activation and repression performance.
- Logic Gene Circuits: Implemented a Toggle Switch and an iFFL to demonstrate programmable gene regulation in V. natriegens.
- Metabolic Engineering Validation: Successfully synthesized naringenin and violacein in V. natriegens, verifying the toolkit's feasibility.
- Computational Modeling: Integrated modeling throughout the project to guide and validate experimental decisions at every stage.
