Integrating CRISPR Genomics with Chemical Genetics to Study Protein-coding Stucture–Function Relationships in Gene Regulation

Abstract

The advent of high-throughput and cost-effective DNA sequencing, alongside revolutionary advances in genomic editing, have democratized genome-scale methods for understanding gene regulation. One fundamental research question for the field—and this dissertation—is then: how do we configure new approaches and wield these increasingly large datasets effectively? In this dissertation, I detail efforts to innovate tiling CRISPR mutagenesis through intentional design of setups and readouts. In Chapter 2, I design a scalable single-cell chemical genomics platform to systematically interrogate agonism and antagonism in AR. I use this framework to not only molecularly define the immediate downstream effects of these processes but also to examine the mechanism of therapeutic candidates in clinical trials. In Chapter 3, I show how CRISPR mutational scanning in combination with drug treatment revealed previously uncharacterized allosteric autoinhibitory mechanism in LSD1 N-terminus. Together, these results demonstrate the potential of high-throughput, structure-agnostic methods in uncovering novel biological insights.