Towards CRISPR Interference Compensators for Single and Multi-Operon Genomic Control in Microbes

This project explores the integration of CRISPRi and remodeling of local supercoiling density as a generalized strategy for engineering the dynamics of the transcriptome in canonical and emerging microbial hosts. Our goal is to develop a rational, quantitative approach for engineering simultaneous control of multiple genes within an operon or multiple operons within a contiguous region of the genome of a new organism. Our approach centers on the fundamental biophysics of DNA supercoiling, using a combination of mathematical modeling, in situ experimental measurements, and genetic engineering to develop CRISPRi circuits that simultaneously control multiple operons or genes within the genome of a target microbe. The biotechnology developed in this project lays the foundation for genome-wide control in an expanding palette of microbial hosts for synthetic biology, with applications in biocomputing, biosensing, and biomaterials.