Nucleic acid detection with CRISPR-Cas13a/C2c2

Nucleic acid detection with CRISPR-Cas13a/C2c2

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 "collater...

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Bibliographic Details
Main Authors: 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, Gootenberg, Jonathan S, Dy, Aaron James, Joung, Julia, Daringer, Nichole Marie, Regev, Aviv, Hung, Deborah T, Collins, James J., Zhang, Feng, Abudayyeh, Omar O.
Other Authors: Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Format: Article
Published: American Association for the Advancement of Science (AAAS) 2017
Online Access:http://hdl.handle.net/1721.1/112745
https://orcid.org/0000-0002-7979-3220
https://orcid.org/0000-0003-0319-5416
https://orcid.org/0000-0001-6656-5002
https://orcid.org/0000-0002-2026-8130
https://orcid.org/0000-0001-8567-2049
https://orcid.org/0000-0002-5560-8246
https://orcid.org/0000-0003-2782-2509
Description
Summary: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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