| Literature DB >> 28408723 |
Jonathan S Gootenberg1,2,3,4,5, Omar O Abudayyeh1,2,3,4,6, Jeong Wook Lee7, Patrick Essletzbichler1,2,3,4, Aaron J Dy1,4,8, Julia Joung1,2,3,4, Vanessa Verdine1,2,3,4, Nina Donghia7, Nichole M Daringer8, Catherine A Freije1,9, Cameron Myhrvold1,9, Roby P Bhattacharyya1, Jonathan Livny1, Aviv Regev1,10, Eugene V Koonin11, Deborah T Hung1, Pardis C Sabeti1,9,12,13, James J Collins14,4,6,7,8, Feng Zhang14,2,3,4.
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.Entities:
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Year: 2017 PMID： 28408723 PMCID： PMC5526198 DOI： 10.1126/science.aam9321
Source DB: PubMed Journal: Science ISSN： 0036-8075 Impact factor: 47.728