I-Corps: Translation potential of an engineered strain of Vibrio natriegens as a chassis for low resource, scalable molecular biology

This I-Corps project is based on the development of an engineered strain of a marine bacterium, Vibrio natriegens, that has been developed to serve as a host organism to scale biotechnology and bioproduction. Microbes that are commonly used today as host organisms for biotechnology and bioproduction are chemically competent, meaning they are chemically treated to take in DNA.

Though commonplace, this process makes cells fragile, requiring investments in equipment and infrastructure to utilize them. In contrast, Vibrio natriegens is naturally competent, meaning it will take in DNA without any chemical intervention. Vibrio natriegens is the fastest growing organism on Earth and does not cause disease.

This bacterial strain has been engineered to enhance scalability and ease of handling to allow for integration with fully automated systems. This scalability and each of handling may accelerate modification and testing and improve protein biomanufacturing as well as the ability to synthesize proteins of interest. These capabilities may shorten the Design-Build-Test-Learn cycle and improve the speed and simplicity of workflows.

With increased speed in these processes, the time and cost of pharmaceutical and vaccine development may be reduced, and new biological solutions to current problems may be developed more rapidly and easily.

This I-Corps project utilizes experiential learning coupled with first-hand investigation of the industry ecosystem to assess the translation potential of an engineered strain of Vibrio natriegens as a chassis for scalable molecular biology and bioproduction. Vibrio natriegens expresses a high rate of natural competency, meaning it will organically take in DNA without chemical or other treatment.

This is different from currently used E. coli strains that must be chemically treated every time DNA is introduced into the cell. Utilizing a genomic insertion of the TfoX master competency regulator gene combined with a specifically designed growth media, a strain of Vibrio natriegens was developed that can be transformed with DNA without the need for special equipment, which means transformation can be completed at room temperature with no heat shock, shaking, media transfers, or incubation.

This technology may lower the cost of synthetic biology by several orders of magnitude and decrease the hands-on time to single minutes. In addition, due to Vibrio natriegens rapid growth rate, an experiment starting with DNA isolation to production of single colonies may be completed in a single day. Recombinant proteins can be rapidly tested and automated at large scales with ease.

Further, cells have been shown to remain competent after transformation, allowing for repeated transformations of the same bacteria. This technology may enable more rapid genetic engineering and protein production.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.