Nucleic acid detection with CRISPR-Cas13a/C2c2
Author(s)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; Gootenberg, Jonathan S; Dy, Aaron James; Joung, Julia; Daringer, Nichole Marie; Regev, Aviv; Hung, Deborah T; Collins, James J.; Zhang, Feng; Abudayyeh, Omar O.; ... Show more Show less
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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.
DepartmentInstitute for Medical Engineering and 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
American Association for the Advancement of Science (AAAS)
Gootenberg, Jonathan S. et al. “Nucleic Acid Detection with CRISPR-Cas13a/C2c2.” Science 356, 6336 (April 2017): 438–442 © 2016 American Association for the Advancement of Science
Author's final manuscript