Discipline: Chemistry and Chemical Sciences
Subcategory: Biochemistry (not Cell and Molecular Biology and Genetics)
Session: 1
Room: Exhibit Hall
Mary Katherine Desrochers - St. Olaf College
Co-Author(s): Chase Weidmann, Ph.D., University of Michigan, Ann Arbor, MI
RNA is a biomolecule useful for the creation of proteins, but it can also function on its own in complex folded structures. Both coding and noncoding RNA might serve as potential therapeutic targets. Our goal is to probe the small molecule binding landscape of RNAs in cells, allowing us to better understand how RNA interactions might be manipulated. Toward this goal, we designed a proof-of-principle strategy for capture and characterization of compound-linked RNAs. Using click chemistry strategies, we covalently linked labeled RNA to protein tags, enabling enrichment by phase separation and read-out by sequencing. RNA-protein complexes were successfully constructed at low rates, and preliminary sequencing results promisingly displayed increased mutation frequency at expected crosslinked nucleotides. With a streamlined way to enrich and locate sites of RNA-protein crosslinks, fragment libraries can be designed to probe all RNAs directly in cells. Before moving to high throughput screening, further optimization is required to achieve higher conversion rates of protein-tagged RNA complexes.
Funder Acknowledgement(s): NSF
Faculty Advisor: Chase Weidmann, cweidman@umich.edu
Role: I worked on the optimization of the click chemistry reaction to aid in the formation of the desired ribonucleoprotein complexes. Additionally, I performed Orthogonal Organic Phase Separation on created complexes and sequenced the resulting RNA to gather preliminary sequencing data.