CRISPR-based molecular diagnostics
CRISPR enzymes have been used to detect nucleic acid. Our lab has worked to develop assays from sample preparation to diagnostic for SARS-Cov-2 (the virus responsible for COVID-19). Our lab has also worked to develop fundamental theories on CRISPR kinetics which govern the limit of detection of these assays.
Cas12a-gRNA complex (5id6)
Nucleic acid detection using fluorescent reporters
Background activity quantification
References
Ashwin Ramachandran and Juan G. Santiago. "CRISPR enzyme kinetics for molecular diagnostics" Analytical Chemistry 93, no. 20 (2021): 7456-7464.
Ashwin Ramachandran, Diego A. Huyke, Eesha Sharma, Malaya K. Sahoo, ChunHong Huang, Niaz Banaei, Benjamin A. Pinsky, and Juan G. Santiago. "Electric field-driven microfluidics for rapid CRISPR-based diagnostics and its application to detection of SARS-CoV-2" Proceedings of the National Academy of Sciences 117, no. 47 (2020): 29518-29525.
Diego A. Huyke, Ashwin Ramachandran, Vladimir I. Bashkirov, Efthalia K. Kotseroglou, Theofilos Kotseroglou, and Juan G. Santiago. "Enzyme Kinetics and Detector Sensitivity Determine Limits of Detection of Amplification-Free CRISPR-Cas12 and Cas13 Diagnostics" Analytical Chemistry 94, no. 27 (2022): 9826-9834.
Charles Blanluet, Diego A. Huyke, Ashwin Ramachandran, Alexandre S. Avaro, and Juan G. Santiago. "Detection and Discrimination of Single Nucleotide Polymorphisms by Quantification of CRISPR-Cas Catalytic Efficiency" Analytical Chemistry 94, no. 43 (2022): 15117-15123.
Alexandre S. Avaro and Juan G. Santiago. "Uncertainty Quantification of Michaelis–Menten Kinetic Rates and Its Application to the Analysis of CRISPR‐Based Diagnostics" Angewandte Chemie International Edition 61, no. 45 (2022): e202209527.
Alexandre S. Avaro and Juan G. Santiago. "A critical review of microfluidic systems for CRISPR assays" Lab Chip 23 (2023): 938-963.