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Nucleic acid detection with CRISPR-Cas13a/C2c2

Jonathan S. GootenbergDepartment of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USAOmar O. AbudayyehDepartment of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USAJeong Wook LeeWyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USAPatrick EssletzbichlerDepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USAAaron J. DyBroad Institute of MIT and Harvard, Cambridge, MA 02142, USAJulia JoungDepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USAVanessa K. VerdineDepartment of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, 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. FreijeCenter for Systems BiologyCameron 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 RegevDepartment of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, 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. SabetiCenter for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, MA 02138, USAJames J. CollinsDepartment of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USAFeng ZhangDepartment of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA

2017en

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Manbada oʻqishwww.ncbi.nlm.nih.gov(yangi oynada ochiladi)

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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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