title: Paired Design of dCas9 as a Systematic Platform for the Detection of Featured Nucleic Acid Sequences in Pathogenic Strains.
abstract: We developed an in vitro DNA detection system using a pair of dCas9 proteins linked to split halves of luciferase. Luminescence was induced upon colocalization of the reporter pair to a ∼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.
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.|Center for Quantitative Biology and Peking-Tsinghua Joint Center for Life Sciences, Peking University , Beijing, 100871, China.
authors.forename: Yihao
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Biology and Center for Genomics and Systems Biology, New York University , 12 Waverly Place, New York, New York 10003, United States.
authors.forename: Long
authors.initials: L
authors.lastname: Qian
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.|CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences , Beijing, 100101, China.|University of Chinese Academy of Science , Beijing, 100149, China.
authors.forename: Weijia
authors.initials: W
authors.lastname: Wei
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Beining
authors.initials: B
authors.lastname: Wang
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Lin
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Wenchao
authors.initials: W
authors.lastname: Liu
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Luze
authors.initials: L
authors.lastname: Xu
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Dongming
authors.initials: D
authors.lastname: Liu
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Sida
authors.initials: S
authors.lastname: Cheng
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Jiaofeng
authors.initials: J
authors.lastname: Li
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Yixuan
authors.initials: Y
authors.lastname: Ye
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Hang
authors.initials: H
authors.lastname: Li
authors.affiliation: Peking University Team for the International Genetically Engineered Machine Competition (iGEM) , Beijing, 100871, China.
authors.forename: Xiaohan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Center for Quantitative Biology and Peking-Tsinghua Joint Center for Life Sciences, Peking University , Beijing, 100871, China.
authors.forename: Yiming
authors.initials: Y
authors.lastname: Dong
authors.affiliation: CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences , Beijing, 100101, China.
authors.forename: Xuejin
authors.initials: X
authors.lastname: Zhao
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences , Beijing, 100101, China.
authors.forename: Cuihua
authors.initials: C
authors.lastname: Liu
authors.affiliation: Center for Quantitative Biology and Peking-Tsinghua Joint Center for Life Sciences, Peking University , Beijing, 100871, China.
authors.forename: Haoqian M
authors.initials: HM
authors.lastname: Zhang
authors.affiliation: Center for Quantitative Biology and Peking-Tsinghua Joint Center for Life Sciences, Peking University , Beijing, 100871, China.|The State Key Laboratory for Artificial Microstructures and Mesoscopic Physics, School of Physics, Peking University , Beijing, 100871, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Ouyang
authors.affiliation: CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences , Beijing, 100101, China.|University of Chinese Academy of Science , Beijing, 100149, China.
authors.forename: Chunbo
authors.initials: C
authors.lastname: Lou
mesh_terms: D001483:Base Sequence / Q000235:genetics*; D064113:CRISPR-Cas Systems / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004720:Endonucleases / Q000235:genetics*; D016680:Genome, Bacterial / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D049449:Luminescence; D009169:Mycobacterium tuberculosis / Q000235:genetics; D009696:Nucleic Acids / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D009696:Nucleic Acids; D008156:Luciferases; D004720:Endonucleases
keywords: DNA detection; dCas9; in vitro pathogenic detection; paired design; split luciferase; tuberculosis