A CRISPR-Cas12a-derived biosensing platform for the highly sensitive detection of diverse small molecules

Mindong Liang, Zilong Li, Weishan Wang, Jiakun Liu, Leshi Liu, Guoliang Zhu, Loganathan Karthik, Man Wang, Ke Feng Wang, Zhong Wang, Jing Yu, Yuting Shuai, Jiaming Yu, Lu Zhang, Zhiheng Yang, Chuan Li, Qian Zhang, Tong Shi, Liming Zhou, Feng XieHuanqin Dai, Xueting Liu, Jingyu Zhang, Guang Liu, Ying Zhuo, Buchang Zhang, Chenli Liu, Shanshan Li, Xuekui Xia, Yaojun Tong, Yanwen Liu, Gil Alterovitz, Gao Yi Tan, Li Xin Zhang

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Besides genome editing, CRISPR-Cas12a has recently been used for DNA detection applications with attomolar sensitivity but, to our knowledge, it has not been used for the detection of small molecules. Bacterial allosteric transcription factors (aTFs) have evolved to sense and respond sensitively to a variety of small molecules to benefit bacterial survival. By combining the single-stranded DNA cleavage ability of CRISPR-Cas12a and the competitive binding activities of aTFs for small molecules and double-stranded DNA, here we develop a simple, supersensitive, fast and high-throughput platform for the detection of small molecules, designated CaT-SMelor (CRISPR-Cas12a- and aTF-mediated small molecule detector). CaT-SMelor is successfully evaluated by detecting nanomolar levels of various small molecules, including uric acid and p-hydroxybenzoic acid among their structurally similar analogues. We also demonstrate that our CaT-SMelor directly measured the uric acid concentration in clinical human blood samples, indicating a great potential of CaT-SMelor in the detection of small molecules.
Original languageEnglish
Article number3672
JournalNature Communications
Issue number1
Publication statusPublished - 2019


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