APPLICATIONS OF TECHNOLOGY:
- Microbiome engineering for medicine, agriculture, biomanufacturing and renewable energy
- Pharmaceutical synthesis
- Commodity chemical and biofuel production
- Livestock research and development
- Enables high-throughput, stable, accurate, efficient gene integration and expression of very large DNA constructs (e.g., pathways)
- Applicable for broad range of bacteria hosts
- Lower-priced approach can be automated
Berkeley Lab researchers including Yasuo Yoshikuni, Gaoyan Wang, Zhiying Zhao, and Jan-Fang Cheng have developed a strategy — Chassis-independent, Recombinase-assisted, Genome Engineering (CRAGE) — to chromosomally integrate and express DNA constructs comprising single genes to complex pathways in a broad range of bacterial hosts. By comparing traditional product and pathway specific modifications to host strains in a high-throughput format, the Berkeley Lab technology reduces the time and cost of identifying candidate production strains in a significant way. Strain-to-strain variation can be minimized due to the use of a pre-defined chromosomal integration site. This development is a major technical step forward in the wholesale transfer of heterologous biosynthetic pathways to bacterial organisms derived from various environments across human, livestock, and plant microbiomes. The technology has been demonstrated to work across a wide variety of bacterial hosts.
The Berkeley Lab technology allows for rapid and efficient transfer and integration of biosynthetic pathway-encoding gene clusters of up to 50 kb and is applicable across many bacterial hosts. Other methods exist to do this, such as the use of plasmid based expression, homologous recombination, or transposition-based chromosomal integration, but at much lower efficiencies and lacking consistent, site-specific integration for relevant comparison across transformants and various bacterial strains.
DEVELOPMENT STAGE: Proven principle
STATUS: Published PCT patent application, PCT/US2017/014788 (WO 2017/176347). Available for licensing or collaborative research.
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