Tools for Genetic Control and Viral Delivery of Multi-Part Gene Cassettes

Register in advance for this meeting:
https://ucsd.zoom.us/meeting/register/tJYsduyhrDsvGt0OlE1PH2g3V1DHtTjaFqSb

 

Friday, May 15, 2020 -
2:00pm to 3:00pm
Zoom
Phillip Kyriakakis

 

Assistant Project Scientist in the Neural Interaction Lab with Professor Todd Coleman

University of California, San Diego

Tools for Genetic Control and Viral Delivery of Multi-Part Gene Cassettes

Abstract: 

Controlling genes with light is a powerful tool for studying biology, and a system that is both reversible and can penetrate tissues is optimal for spatiotemporal and in vivo gene control. The highly sensitive red/far-red light “PhytochromeB gene switch” has those characteristics. Yet past attempts to genetically encode this system in non-plant eukaryotic cells have been limited by the inability to produce the necessary chromophore. We found that matching the species of the Ferredoxin enzyme system with the metabolic pathway is required for electron transport to synthesize the chromophore efficiently, enabling us to genetically encode the entire system in mammalian cells (Kyriakakis et al., 2018)-(WO2017192920A1). This species matching concept is now being used by many others to engineer metabolic pathways to produce fertilizer in yeast and in crops (Allen et al., 2017) (Burén and Rubio, 2018).  Because of the large size of the genes encoding these metabolic pathways, my current efforts focus on developing tools for efficient DNA assembly and gene delivery. Using a new method that allows for recursive assembly of DNA, uLoop, we are constructing a library of parts (promoters, PolyA, optogenetics, fluorescent reporters, etc.) to assemble genetic systems quickly. To deliver these large constructs in vivo, we are also developing several nontoxic viral vectors to be compatible with our uLoop DNA parts. Combining uLoop assembly with large payload viral vectors and optogenetics, we are building tools to study development/disease in mouse models of hearing loss and for neural circuit tracing.

Bio: 

Dr. Phillip Kyriakakis is an Assistant Project Scientist in the Neural Interaction Lab with Professor Todd Coleman here at UC San Diego. Phillip Kyriakakis received his BS in Biochemistry in 2008 from the University of Massachusetts at Boston, where he worked in Dr. Alexey Veraksa’s proteomics/developmental biology lab. In 2014, Dr. Kyriakakis received his PhD in Biology from UCSD, with a specialization in Multiscale Biology, where he identified and characterized a metabolic enzyme, glucokinase, that forms actin-like filaments. As a postdoctoral fellow in Bioengineering with Todd Coleman, Dr. Kyriakakis has developed (plant) PhyB optogenetic tools that can be genetically encoded in animal cells. This allows for the control of a gene using red and far-red light to induce and halt gene activity in mammalian cells. His work now focuses on building tools and platforms for synthetic biological or biochemical systems.