Nucleic acid detection with CRISPR-Cas13a/C2c2

Jonathan S. Gootenberg; Omar O. Abudayyeh; Jeong Wook Lee; Patrick Essletzbichler; Aaron J. Dy; Julia Joung; Vanessa Verdine; Nina Donghia; Nichole M. Daringer; Catherine A. Freije; Cameron Myhrvold; Roby P. Bhattacharyya; Jonathan Livny; Aviv Regev; Eugene V. Koonin; Deborah T. Hung; Pardis C. Sabeti; James J. Collins; Feng Zhang

doi:10.1126/science.aam9321

Nucleic acid detection with CRISPR-Cas13a/C2c2

Jonathan S. Gootenberg, Omar O. Abudayyeh, Jeong Wook Lee, Patrick Essletzbichler, Aaron J. Dy, Julia Joung, Vanessa Verdine, Nina Donghia, Nichole M. Daringer, Catherine A. Freije, Cameron Myhrvold, Roby P. Bhattacharyya, Jonathan Livny, Aviv Regev, Eugene V. Koonin, Deborah T. Hung, Pardis C. Sabeti, James J. Collins, Feng Zhang

Research output: Contribution to journal › Article › peer-review

2659 Scopus citations

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.

Original language	English (US)
Pages (from-to)	438-442
Number of pages	5
Journal	Science
Volume	356
Issue number	6336
DOIs	https://doi.org/10.1126/science.aam9321
State	Published - Apr 28 2017
Externally published	Yes

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Gootenberg, J. S., Abudayyeh, O. O., Lee, J. W., Essletzbichler, P., Dy, A. J., Joung, J., Verdine, V., Donghia, N., Daringer, N. M., Freije, C. A., Myhrvold, C., Bhattacharyya, R. P., Livny, J., Regev, A., Koonin, E. V., Hung, D. T., Sabeti, P. C., Collins, J. J., & Zhang, F. (2017). Nucleic acid detection with CRISPR-Cas13a/C2c2. Science, 356(6336), 438-442. https://doi.org/10.1126/science.aam9321

@article{b662d4dabc2c4e038e09559b641d5f8f,

title = "Nucleic acid detection with CRISPR-Cas13a/C2c2",

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.",

author = "Gootenberg, {Jonathan S.} and Abudayyeh, {Omar O.} and Lee, {Jeong Wook} and Patrick Essletzbichler and Dy, {Aaron J.} and Julia Joung and Vanessa Verdine and Nina Donghia and Daringer, {Nichole M.} and Freije, {Catherine A.} and Cameron Myhrvold and Bhattacharyya, {Roby P.} and Jonathan Livny and Aviv Regev and Koonin, {Eugene V.} and Hung, {Deborah T.} and Sabeti, {Pardis C.} and Collins, {James J.} and Feng Zhang",

year = "2017",

month = apr,

day = "28",

doi = "10.1126/science.aam9321",

language = "English (US)",

volume = "356",

pages = "438--442",

journal = "Science",

issn = "0036-8075",

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Gootenberg, JS, Abudayyeh, OO, Lee, JW, Essletzbichler, P, Dy, AJ, Joung, J, Verdine, V, Donghia, N, Daringer, NM, Freije, CA, Myhrvold, C, Bhattacharyya, RP, Livny, J, Regev, A, Koonin, EV, Hung, DT, Sabeti, PC, Collins, JJ & Zhang, F 2017, 'Nucleic acid detection with CRISPR-Cas13a/C2c2', Science, vol. 356, no. 6336, pp. 438-442. https://doi.org/10.1126/science.aam9321

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

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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DO - 10.1126/science.aam9321

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AN - SCOPUS:85017652697

SN - 0036-8075

VL - 356

SP - 438

EP - 442

JO - Science

JF - Science

IS - 6336

ER -

Nucleic acid detection with CRISPR-Cas13a/C2c2

Abstract

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