TY - JOUR
T1 - Nucleic acid detection with CRISPR-Cas13a/C2c2
AU - Gootenberg, Jonathan S.
AU - Abudayyeh, Omar O.
AU - Lee, Jeong Wook
AU - Essletzbichler, Patrick
AU - Dy, Aaron J.
AU - Joung, Julia
AU - Verdine, Vanessa
AU - Donghia, Nina
AU - Daringer, Nichole M.
AU - Freije, Catherine A.
AU - Myhrvold, Cameron
AU - Bhattacharyya, Roby P.
AU - Livny, Jonathan
AU - Regev, Aviv
AU - Koonin, Eugene V.
AU - Hung, Deborah T.
AU - Sabeti, Pardis C.
AU - Collins, James J.
AU - Zhang, Feng
N1 - Publisher Copyright:
Copyright © 2016 by the American Association for the Advancement of Science; All rights reserved.
PY - 2017/4/28
Y1 - 2017/4/28
N2 - 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.
AB - 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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U2 - 10.1126/science.aam9321
DO - 10.1126/science.aam9321
M3 - Article
C2 - 28408723
AN - SCOPUS:85017652697
SN - 0036-8075
VL - 356
SP - 438
EP - 442
JO - Science
JF - Science
IS - 6336
ER -