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Twist Bioscience HQ
681 Gateway Blvd
South San Francisco, CA 94080
Pathway Engineering Through 5kb Gene Synthesis
Cloned genes up to 5kb can be used to encode entire pathways to trivialize the build step of the design-build-test cycle.
The design-build-test cycle is vital to metabolic engineering applications in which researchers re-program microorganisms to produce non-native molecules of interest. However, reprogramming cells to synthesize molecules in amounts that are sufficient for purification often requires many iterations to achieve viable titers. Innovations affording a rapid design-build-test cycle are therefore highly sought after. As a proof-of-concept accelerated workflow for high throughput multi-gene testing, we applied the Twist oligonucleotide synthesis platform to synthesize genes as long as 5 kb. Multiple genes involved in carotenoid synthesis pathways were cloned into a single plasmid, and valuable pigment molecules could be rapidly produced.
Covered in this Application Note
Design of a carotenoid synthesis pathway for an E. coli chassis.
A trivial build of the pathway using Twist's ordering and synthesis platform
Testing of Carotenoid synthesis.
Optimization of the pathway to produce valuable pigment molecules.
