February 2020 Kentucky Lake Section Meeting
Dry Ground Brewery
3121 Broadway St (Next to Mellow Mushroom)
Paducah, KY 24003
Thursday, February 27th
Dinner 6:00 pm
Presentation 7:00 pm
Dinner: Pizza from Mellow Mushroom ($10, $5 for students)
Program: Controlling CRISPR-Cas Enzymes with Chemically Modified Nucleic Acids
Dr. Keith T. Gagnon, Southern Illinois University
Abstract: Clustered regularly interspaced short palindromic repeat (CRISPR) RNAs (crRNAs) and their associated effector (Cas) enzymes have revolutionized basic biotechnology and are a promising technology for treating genetic diseases. However, CRISPR-Cas enzymes have some unwanted properties, such as off-target gene editing, and are not ideal for drug development. To better understand the role of the crRNA component and the requirements for making CRISPR-Cas enzymes more drug-like, we have explored a variety of chemical modifications to the crRNA. We uncovered rules for modification, such as A-form-like helical structure, flexibility, and moieties with low bulkiness as keys to chemical compatibility. We also identified several critical positions that seem to require a 2′ hydroxyl on the RNA ribose ring and are working to understand this dependency and ultimately remove all labile RNA nucleotides from the crRNA. Using chemically modified nucleic acids, we have also created potent inhibitors of the CRISPR-Cas9 enzyme. These small nucleic acid-based inhibitors (SNuBs) bind with very high affinity and can block gene editing inside of cells. CRISPR SNuBs are being further optimized for even greater potency and testing in animal models of gene editing. Together, these studies have improved our understanding of enzyme activity, identified ways to control CRISPR-Cas enzyme activity, and are contributing to safe and practical development of CRISPR as a human therapeutic.
Speaker Bio: Dr. Gagnon is currently an Assistant Professor in both the Department of Chemistry & Biochemistry AND the Department of Biochemistry & Molecular Biology in the School of Medicine at Southern Illinois University in Carbondale, IL. He also serves as the founder and CEO of Guide BioSci, Inc.. Dr. Gagnon received his B.S. and Ph.D. (in Biochemistry) from North Carolina State University—his graduate work being completed under the supervision of Dr. E. Stuart Maxwell—and, he completed a post doc with Dr. David R. Corey at the UT Southwestern Medical Center. Among Dr. Gagnon’s many awards, he has been recognized with the Outstanding Young Alumni Award from North Carolina State University in 2016 and the Oligonucleotide Therapeutics Society Outstanding Young Investigator Award in 2017.