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A Step-by-Step Protocol for COMPASS, a Synthetic Biology Tool for Combinatorial Gene Assembly
Gita Naseri1, Bernd Mueller-Roeber2,3,4
1Department of Molecular Biology, University of Potsdam, Potsdam, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|August 19, 2020
Summary
Eliminating metabolic flux imbalances is key for industrial chemical production. The COMbinatorial Pathway ASSembly (COMPASS) method rapidly generates balanced gene expression constructs for microbial cell factories.
Area of Science:
- Synthetic biology
- Metabolic engineering
- Molecular biology
Background:
- Industrial chemical production relies on microbial cell factories.
- Metabolic flux imbalances hinder efficient production.
- Optimal genetic pathway design is often unknown beforehand.
Purpose of the Study:
- To develop a rapid cloning method for balanced expression of multiple genes in biochemical pathways.
- To enable high-throughput generation of DNA constructs for pathway engineering.
- To facilitate the optimization of microbial cell factories for chemical production.
Main Methods:
- COMbinatorial Pathway ASSembly (COMPASS) uses inducible plant transcription factors in yeast.
- Homologous recombination is employed for modular parts assembly.
- CRISPR/Cas9 facilitates one-step multilocus gene integration.
Main Results:
- COMPASS generates thousands of DNA constructs in a modular, parallel, high-throughput manner.
- A positive selection scheme identifies correctly assembled pathway variants.
- The method enables balanced gene expression for improved pathway function.
Conclusions:
- COMPASS is an effective method for balancing gene expression in microbial systems.
- This approach accelerates the design and construction of synthetic pathways.
- COMPASS has broad applications in synthetic biology and metabolic engineering.
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