BME PhD Prospectus Defense - James DiCarlo

9:30 am on Tuesday, August 13, 2013
44 Cummington St, Room 401
Title: Multiplex Genome Engineering in Saccharomyces cerevisiae

Jim Collins (BME co-advisor, Chair)
Wilson Wong
Ahmad Khalil
George Church (Research Advisor)

High-frequency multiplexed genome engineering techniques allow for rapid directed genome modification and have been developed in several organisms in recent years. High-throughput generation of multiple alleles allow for investigation of epistatic interactions between DNA regulatory regions and proteins. Additionally, the introduction of diversity in a targeted fashion can be used to optimize biosynthetic pathways. This project aims to develop a method for high-frequency multiplexed genome engineering in the industrially and academically important model eukaryotic organism, Saccharomyces cerevisiae. To develop this method, two approaches will be explored. The first approach involves the manipulation of the endogenous homologous recombination system in yeast, while the second approach examines the employment of programmable endonucleases to stimulate and select for recombination events. As a proof of concept for the optimum method developed, industrially relevant phenotypes of yeast as well as a biosynthetic pathway will be generated using the protocol.