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Nucleic acid detection with CRISPR-Cas13a/C2c2

Jonathan S. GootenbergBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAOmar O. AbudayyehBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAJeong Wook LeeWyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USAPatrick EssletzbichlerBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAAaron J. DyBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAJulia JoungBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAVanessa K. VerdineBroad Institute of MIT and Harvard, Cambridge, MA 02142, USANina M. DonghiaWyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USANichole M. DaringerInstitute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USACatherine A. FreijeBroad Institute of MIT and Harvard, Cambridge, MA 02142, USACameron MyhrvoldBroad Institute of MIT and Harvard, Cambridge, MA 02142, USARoby P. BhattacharyyaBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAJonathan LivnyBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAAviv RegevBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAEugene V. KooninNational Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, MD 20894, USADeborah T. HungBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAPardis C. SabetiBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAJames J. CollinsBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAFeng ZhangBroad Institute of MIT and Harvard, Cambridge, MA 02142, USA

2017en

ABI

Annotatsiya

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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DOI: 10.1126/science.aam9321

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