| Literature DB >> 31413315 |
Mindong Liang1,2, Zilong Li3, Weishan Wang3, Jiakun Liu1,4, Leshi Liu1, Guoliang Zhu1, Loganathan Karthik1, Man Wang5, Ke-Feng Wang1, Zhong Wang6, Jing Yu7, Yuting Shuai1, Jiaming Yu1, Lu Zhang1, Zhiheng Yang1, Chuan Li1, Qian Zhang1, Tong Shi1, Liming Zhou1, Feng Xie3, Huanqin Dai3, Xueting Liu1, Jingyu Zhang1, Guang Liu1, Ying Zhuo1, Buchang Zhang2, Chenli Liu4, Shanshan Li8, Xuekui Xia9, Yaojun Tong10, Yanwen Liu11, Gil Alterovitz12, Gao-Yi Tan13, Li-Xin Zhang14.
Abstract
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.Entities:
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Year: 2019 PMID： 31413315 PMCID： PMC6694116 DOI： 10.1038/s41467-019-11648-1
Source DB: PubMed Journal: Nat Commun ISSN： 2041-1723 Impact factor: 14.919