Session 8, Abstract 46
ENGINEERING IMPROVED HOST STRAINS FOR A SYNTHETIC GENETIC OSCILLATOR
Joy Doong* and Justin C. Hardin (Andrew Yao and Marc T. Facciotti), Dept. of Biomedical Engineering and Genome Center, University of California, Davis, 1 Shields Ave, Davis, CA 95616
A genetic oscillator is a genetic circuit that switches between two states with a consistent period. This could act as a timekeeper or counter in complex biological devices. Previously published oscillator circuits have delivered inconsistent and unsustained oscillating outputs. We hypothesize that improvements can be made by reducing the influence of two factors: (1) the potential for non-linear inducible response to external stimuli and (2) the molecular crosstalk between the oscillator circuits and the related pathways in the host. To address these factors, we are building new genomically engineered host strains by deleting specific genes using the Scarless Cas9 Assisted Recombineering method, which we have successfully adapted and refined to make single modifications in about two weeks. Thus far, we have made two of the four planned modifications, verified by either genetic or functional screens. We will sequence genomes to verify our modifications, and then test the knockout strains by examining differences in the input-output behavior of promoters in response to exogenous inducers. Finally we will study how the new host strains influence the behavior of the oscillator circuits.