1. Nucleic acid detection with CRISPR-Cas13a/C2c2
- Author
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Massachusetts Institute of Technology. Institute for Medical Engineering & Science, Harvard University--MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences, McGovern Institute for Brain Research at MIT, Gootenberg, Jonathan S, Abudayyeh, Omar Osama, Dy, Aaron James, Joung, Julia, Daringer, Nichole Marie, Regev, Aviv, Hung, Deborah T, Collins, James J., Zhang, Feng, Lee, Jeong Wook, Essletzbichler, Patrick, Verdine, Vanessa, Donghia, Nina, Freije, Catherine A., Myhrvold, Cameron, Bhattacharyya, Roby P., Livny, Jonathan, Koonin, Eugene V., Pardis, C. Sabeti, Abudayyeh, Omar O., Massachusetts Institute of Technology. Institute for Medical Engineering & Science, Harvard University--MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences, McGovern Institute for Brain Research at MIT, Gootenberg, Jonathan S, Abudayyeh, Omar Osama, Dy, Aaron James, Joung, Julia, Daringer, Nichole Marie, Regev, Aviv, Hung, Deborah T, Collins, James J., Zhang, Feng, Lee, Jeong Wook, Essletzbichler, Patrick, Verdine, Vanessa, Donghia, Nina, Freije, Catherine A., Myhrvold, Cameron, Bhattacharyya, Roby P., Livny, Jonathan, Koonin, Eugene V., Pardis, C. Sabeti, and Abudayyeh, Omar O.
- Abstract
Rapid, inexpensive, and sensitive nucleic acid detection may aid point-of-care pathogen detection, genotyping, and disease monitoring. The RNA-guided, RNA-targeting clustered regularly interspaced short palindromic repeats (CRISPR) effector Cas13a (previously known as C2c2) exhibits a "collateral effect" of promiscuous ribonuclease activity upon target recognition. We combine the collateral effect of Cas13a with isothermal amplification to establish a CRISPR-based diagnostic (CRISPR-Dx), providing rapid DNA or RNA detection with attomolar sensitivity and single-base mismatch specificity. We use this Cas13a-based molecular detection platform, termed Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK), to detect specific strains of Zika and Dengue virus, distinguish pathogenic bacteria, genotype human DNA, and identify mutations in cell-free tumor DNA. Furthermore, SHERLOCK reaction reagents can be lyophilized for cold-chain independence and long-term storage and be readily reconstituted on paper for field applications., United States. Air Force Office of Scientific Research (Grant FA9550-14-1-0060), Defense Threat Reduction Agency (DTRA) (Grant HDTRA1-14-1-0006), National Institute of Mental Health (U.S.) (Grant 5DP1-MH100706), National Institutes of Health (U.S.) (Grant 1R01-MH110049)
- Published
- 2017