Nucleic acid detection with CRISPR-Cas13a/C2c2
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nihms873520.pdf
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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
Date Issued
April 2017
Journal
Science
Publisher
American Association for the Advancement of Science (AAAS)
Citation
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
Version
Author's final manuscript
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.
MIT Department
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
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DOI of Published Version
https://doi.org/10.1126/SCIENCE.AAM9321
