(635g) Engineered CRISPR/Cas Systems As Diagnostics and Therapeutics (INVITED SPEAKER)
AIChE Annual Meeting
2020
2020 Virtual AIChE Annual Meeting
Food, Pharmaceutical & Bioengineering Division
Integrative Approaches to Study Disease Mechanisms and Therapies
Friday, November 20, 2020 - 9:30am to 9:45am
The Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated (CRISPR/Cas) systems have emerged as potent and versatile genome editing tools with a wide range of applications including understanding disease mechanisms1-2. However, clinical translation of CRISPR/Cas systems as diagnostics and/or therapeutics heavily relies on: 1) improving the efficiency and specificity of CRISPR/Cas systems, 2) developing targeted delivery strategies, and 3) engineering the ability to control their activity on demand3. Towards these goals, we previously created a light-activatable CRISPR-PLUS technology that allowed spatiotemporal control of gene editing in vitro5. In parallel, we also developed a peptide-based CRISPR-GPS system for targeted delivery of CRISPR/Cas components into a variety of cell lines6 and in a xenograft model of human ovarian cancer in mice.
Our lab recently engineered a CRISPR-ENHACE technology by modifying CRISPR RNA (crRNAs) and optimizing conditions that allowed unprecedented enhancement in the sensitivity of CRISPR/Cas12a for nucleic acid detection with improved specificity. While engineering CRISPR-ENHANCE, we discovered a mechanism by which a Cas12a enhances the rate of enzymatic cleavage by 3.5-fold, making it one of the fastest known CRISPR/Cas systems in terms of collateral cleavage activity. We further applied the CRISPR-ENHANCE system with isothermal amplification strategies for detecting prostate cancer in the urine as well as for developing rapid diagnostics for HIV, HCV, and SARS-CoV-27. In this talk, the fundamentals of engineered CRISPR/Cas technologies in Figure 1 will be introduced with a focus on the development of a COVID-19 testing kit using CRISPR-ENHANCE technology.
References:
[1] M. Jinek, et al., Science, 2012, 337, 816-821.
[2] B. Zetsche, et al., Cell, 2015, 163, 759-771.
[3] Komor, et al., Cell, 2017, 169 (3), 559.
[4] S. Q. Tsai, Nat. Biotech., 2015, 33, 187–197.
[5] P.K. Jain, et al., Angew. Chem., 2016, 55, 12440.
[6] P.K. Jain, et al., Nanoscale, 2019, 11, 21317-21323.
[7] L. Nguyen, et al., BioRxiv, 2020, 10.1101/2020.04.13.036079.