Journal
SCIENCE
Volume 356, Issue 6336, Pages 438-+Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.aam9321
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Funding
- Paul and Daisy Soros Fellowship
- National Defense Science and Engineering Fellowship
- U.S. Department of Energy Computational Science Graduate Fellowship
- NIH through a National Institute of Allergies and Infectious Diseases grant [R01AI117043]
- NSF Graduate Research Fellowship
- Air Force Office of Scientific Research grant [FA9550-14-1-0060]
- M. and L. Benioff
- J. and A. Bekenstein
- Howard Hughes Medical Institute
- Defense Threat Reduction Agency grant [HDTRA1-14-1-0006]
- Paul G. Allen Frontiers Group
- Wyss Institute
- NIH through National Institute of Mental Health grants [5DP1-MH100706, 1R01-MH110049]
- NSF
- New York Stem Cell
- Simons Foundation
- Vallee Foundation
- Paul G. Allen Family Foundation
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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.
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