1. Paired Design of dCas9 as a Systematic Platform for the Detection of Featured Nucleic Acid Sequences in Pathogenic Strains
- Author
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Xiaohan Zhang, Xuejin Zhao, Jiaofeng Li, Sida Cheng, Long Qian, Yixuan Ye, Haoqian Zhang, Wenchao Liu, Dongming Liu, Yiming Dong, Chunbo Lou, Yihao Zhang, Yu Wang, Luze Xu, Hang Li, Xiang Li, Cui Hua Liu, Beining Wang, Qi Ouyang, Pingping Lin, and Weijia Wei
- Subjects
DNA, Bacterial ,0301 basic medicine ,Luminescence ,Biomedical Engineering ,Biology ,Sensitivity and Specificity ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Genome ,Paired design ,03 medical and health sciences ,Nucleic Acids ,Luciferase ,Luciferases ,Sequence (medicine) ,Genetics ,Base Sequence ,Colocalization ,Mycobacterium tuberculosis ,General Medicine ,Endonucleases ,Dna detection ,030104 developmental biology ,Nucleic acid ,Mycobacterium tuberculosis DNA ,CRISPR-Cas Systems ,Genome, Bacterial - 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.
- Published
- 2016