Nucleic acid detection with CRISPR-Cas13a/C2c2

被引：2016

作者：

Gootenberg, Jonathan S. ^{[1
,2
,3
,4
,5
]}

Abudayyeh, Omar O. ^{[1
,2
,3
,4
,6
]}

Lee, Jeong Wook ^{[7
]}

Essletzbichler, Patrick ^{[1
,2
,3
,4
]}

Dy, Aaron J. ^{[1
,4
,8
]}

Joung, Julia ^{[1
,2
,3
,4
]}

Verdine, Vanessa ^{[1
,2
,3
,4
]}

Donghia, Nina ^{[7
]}

Daringer, Nichole M. ^{[8
]}

Freije, Catherine A. ^{[1
,9
]}

Myhrvold, Cameron ^{[1
,9
]}

Bhattacharyya, Roby P. ^{[1
]}

Livny, Jonathan ^{[1
]}

Regev, Aviv ^{[1
,10
]}

Koonin, Eugene V. ^{[11
]}

Hung, Deborah T. ^{[1
]}

Sabeti, Pardis C. ^{[1
,9
,12
,13
]}

Collins, James J. ^{[1
,4
,6
,7
,8
]}

Zhang, Feng ^{[1
,2
,3
,4
]}

机构：

[1] Broad Inst & MIT Harvard, Cambridge, MA 02142 USA

[2] MIT, McGovern Inst Brain Res, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[3] MIT, Dept Brain & Cognit Sci, E25-618, Cambridge, MA 02139 USA

[4] MIT, Dept Biol Engn, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[5] Harvard Med Sch, Dept Syst Biol, Boston, MA 02115 USA

[6] MIT, Dept Hlth Sci & Technol, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[7] Harvard Univ, Wyss Inst Biol Inspired Engn, Boston, MA 02115 USA

[8] MIT, Inst Med Engn & Sci, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[9] Harvard Univ, Dept Organism & Evolutionary Biol, Ctr Syst Biol, Cambridge, MA 02138 USA

[10] MIT, Dept Biol, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[11] NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA

[12] Harvard Sch Publ Hlth, Dept Immunol & Infect Dis, Boston, MA 02115 USA

[13] Howard Hughes Med Inst, Chevy Chase, MD 20815 USA

来源：

SCIENCE | 2017年 / 356卷 / 6336期

关键词：

CIRCULATING TUMOR DNA; CRISPR-CAS SYSTEMS; ZIKA VIRUS; AMPLIFICATION; PROTEINS;

D O I：

10.1126/science.aam9321

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：438 / +

页数：5

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