Paired Design of dCas9 as a Systematic Platform for the Detection of Featured Nucleic Acid Sequences in Pathogenic Strains

被引：160

作者：

Zhang, Yihao ^{[1
,2
,3
]}

Qian, Long ^{[5
,6
]}

Wei, Weijia ^{[1
,4
,9
]}

Wang, Yu ^{[1
]}

Wang, Beining ^{[1
]}

Lin, Pingping ^{[1
]}

Liu, Wenchao ^{[1
]}

Xu, Luze ^{[1
]}

Li, Xiang ^{[1
]}

Liu, Dongming ^{[1
]}

Cheng, Sida ^{[1
]}

Li, Jiaofeng ^{[1
]}

Ye, Yixuan ^{[1
]}

Li, Hang ^{[1
]}

Zhang, Xiaohan ^{[1
]}

Dong, Yiming ^{[2
,3
]}

Zhao, Xuejin ^{[4
]}

Liu, Cuihua ^{[7
]}

Zhang, Haoqian M. ^{[2
,3
]}

Ouyang, Qi ^{[2
,3
,8
]}

Lou, Chunbo ^{[4
,9
]}

机构：

[1] Peking Univ, Team Int Genetically Engn Machine Competit iGEM, Beijing 100871, Peoples R China

[2] Peking Univ, Ctr Quantitat Biol, Beijing 100871, Peoples R China

[3] Peking Univ, Peking Tsinghua Joint Ctr Life Sci, Beijing 100871, Peoples R China

[4] Chinese Acad Sci, Inst Microbiol, CAS Key Lab Microbial Physiol & Metab Engn, Beijing 100101, Peoples R China

[5] NYU, Dept Biol, 12 Waverly Pl, New York, NY 10003 USA

[6] NYU, Ctr Genom & Syst Biol, 12 Waverly Pl, New York, NY 10003 USA

[7] Chinese Acad Sci, Inst Microbiol, CAS Key Lab Pathogen Microbiol & Immunol, Beijing 100101, Peoples R China

[8] Peking Univ, Sch Phys, State Key Lab Artificial Microstruct & Mesoscop P, Beijing 100871, Peoples R China

[9] Univ Chinese Acad Sci, Beijing 100149, Peoples R China

来源：

ACS SYNTHETIC BIOLOGY | 2017年 / 6卷 / 02期

基金：

中国国家自然科学基金;

关键词：

dCas9; paired design; split luciferase; tuberculosis; DNA detection; in vitro pathogenic detection; MEDIATED ISOTHERMAL AMPLIFICATION; MOLECULAR BEACONS; SPECIFICITY; CAS9; ENDONUCLEASE;

D O I：

10.1021/acssynbio.6b00215

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We developed an in vitro DNA detection system using a pair of dCas9 proteins linked to split halves of luciferase. Luminescence was induced upon coloealization of the reporter pair to a similar to 44 bp target sequence defined by sgRNAs. We used the system to detect Mycobacterium tuberculosis DNA with high specificity and sensitivity. The reprogrammability of dCas9 was further leveraged in an array design that accesses sequence information across the entire genome.

引用

页码：211 / 216

页数：6

共 21 条

[1] Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease [J].

Anders, Carolin ;

Niewoehner, Ole ;

Duerst, Alessia ;

Jinek, Martin .

NATURE, 2014, 513 (7519) :569-+

[2] Halo-tag mediated self-labeling of fluorescent proteins to molecular beacons for nucleic acid detection [J].

Blackstock, Daniel ;

Chen, Wilfred .

CHEMICAL COMMUNICATIONS, 2014, 50 (89) :13735-13738

[3]

Chavez A, 2015, NAT METHODS, V12, P326, DOI [10.1038/nmeth.3312, 10.1038/NMETH.3312]

[4] Dynamic Imaging of Genomic Loci in Living Human Cells by an Optimized CRISPR/Cas System [J].

Chen, Baohui ;

Gilbert, Luke A. ;

Cimini, Beth A. ;

Schnitzbauer, Joerg ;

Zhang, Wei ;

Li, Gene-Wei ;

Park, Jason ;

Blackburn, Elizabeth H. ;

Weissman, Jonathan S. ;

Qi, Lei S. ;

Huang, Bo .

CELL, 2013, 155 (07) :1479-1491

[5] The new frontier of genome engineering with CRISPR-Cas9 [J].

Doudna, Jennifer A. ;

Charpentier, Emmanuelle .

SCIENCE, 2014, 346 (6213) :1077-+

[6] A Single Molecular Beacon Probe Is Sufficient for the Analysis of Multiple Nucleic Acid Sequences [J].

Gerasimova, Yulia V. ;

Hayson, Aaron ;

Ballantyne, Jack ;

Kolpashchikov, Dmitry M. .

CHEMBIOCHEM, 2010, 11 (12) :1762-1768

[7] CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes [J].

Gilbert, Luke A. ;

Larson, Matthew H. ;

Morsut, Leonardo ;

Liu, Zairan ;

Brar, Gloria A. ;

Torres, Sandra E. ;

Stern-Ginossar, Noam ;

Brandman, Onn ;

Whitehead, Evan H. ;

Doudna, Jennifer A. ;

Lim, Wendell A. ;

Weissman, Jonathan S. ;

Qi, Lei S. .

CELL, 2013, 154 (02) :442-451

[8] Fusion of catalytically inactive Cas9 to Fokl nuclease improves the specificity of genome modification [J].

Guilinger, John P. ;

Thompson, David B. ;

Liu, David R. .

NATURE BIOTECHNOLOGY, 2014, 32 (06) :577-+

[9] A real-time characterization method to rapidly optimize molecular beacon signal for sensitive nucleic acids analysis [J].

Hsieh, Albert Tsung-Hsi ;

Pan, Patrick J. ;

Lee, Abraham P. .

ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2014, 406 (13) :3059-3067

[10] DNA targeting specificity of RNA-guided Cas9 nucleases [J].

Hsu, Patrick D. ;

Scott, David A. ;

Weinstein, Joshua A. ;

Ran, F. Ann ;

Konermann, Silvana ;

Agarwala, Vineeta ;

Li, Yinqing ;

Fine, Eli J. ;

Wu, Xuebing ;

Shalem, Ophir ;

Cradick, Thomas J. ;

Marraffini, Luciano A. ;

Bao, Gang ;

Zhang, Feng .

NATURE BIOTECHNOLOGY, 2013, 31 (09) :827-+

← 1 2 3 →