Team 1
A Continuous Directed Evolution Platform for the Generation of Optogenetic Promoters
Team Members:
Johnpaul Ciccosanti
Margaret Cook
Spencer Hauxhurst
Nikolai Shteinman
Michelle Tieu
Team Mentor:
A Continuous Directed Evolution Platform for the Generation of Optogenetic Promoters
Team Members:
Johnpaul Ciccosanti
Margaret Cook
Spencer Hauxhurst
Nikolai Shteinman
Michelle Tieu
Team Mentor:
Dr. Benjamin Bartelle, PhD - Arizona State University, SBHSE
YouTube Link:
View the video link below before joining the zoom meeting
Zoom Link:
https://asu.zoom.us/j/88572009772
Abstract
Abstract: The vast majority of drugs produced under the Food and Drug Administration (FDA) are classified as biologics, generally defined as engineered proteins. Leveraging synthetic biology to create natural producers of these biologic pharmaceuticals is both sustainable and financially appealing. Industry has adopted these “microbial factories” to produce drugs at a fraction of the cost. In doing so, they have large amounts of recombinant systems that require regulation of gene expression throughout the process. However, the current methods for doing so in an industrial setting are typically chemical in nature and are expensive, difficult, and time-consuming. Optogenetic promoters are activated and inactivated by light alone, providing an effective, inexpensive, and scalable option. There is existing technology that allows for regulation using blue light, but a lack of promoters responding to multiple wavelengths limits the applications, especially for an industrial setting. We propose a directed evolution approach for the generation and optimization of promoters that respond to additional wavelengths. Our device, blueGENES, and the corresponding plasmid kit are adapted for laboratory use to simplify the process of tuning optogenetic promoters to a desired wavelength. The phage-assisted continuous directed evolution system can maintain cell culture conditions at a constant turbidity using a laser sensor, produce light of varying intensities at wavelengths of 520 nm, 490 nm, and 465 nm, and use magnetic fans to control stir bars. Directed evolution of an optogenetic construct indicated a heightened response to the desired wavelength.
