Paired Design of dCas9 as a Systematic Platform for the Detection of Featured Nucleic Acid Sequences in Pathogenic Strains
Yihao Zhang(Peking University), Long Qian(New York University), Weijia Wei(Chinese Academy of Sciences), Yu Wang(Peking University), Beining Wang(Peking University), Pingping Lin(Peking University), Wenchao Liu(Peking University), Luze Xu(Peking University), Xiang Li(Peking University), Dongming Liu(Peking University), Sida Cheng(Peking University), Jiaofeng Li(Peking University), Yixuan Ye(Peking University), Hang Li(Peking University), Xiaohan Zhang(Peking University), Yiming Dong(Peking University), Xuejin Zhao(Chinese Academy of Sciences), Cuihua Liu(Chinese Academy of Sciences), Haoqian Zhang(Peking University), Qi Ouyang(Peking University), Chunbo Lou(Chinese Academy of Sciences)
Cited by 185
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.
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