A CRISPR-Cas12a-derived biosensing platform for the highly sensitive detection of diverse small molecules
A CRISPR-Cas12a-derived biosensing platform for the highly sensitive detection of diverse small molecules
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源自 CRISPR-Cas12a 的生物传感平台，用于高灵敏度检测多种小分子
DOI：
10.1038/s41467-019-11648-1
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发表时间：
2019-08-14
影响因子：
16.6
通讯作者：
Zhang, Li-Xin
中科院分区：
文献类型：
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作者：
Liang, Mindong;Li, Zilong;Zhang, Li-Xin
除了基因组编辑，CRISPR-Cas12a最近还被用于具有全分子敏感性的DNA检测应用，但据我们所知，它还没有用于小分子的检测。细菌变构转录因子(ATF)已经进化为对各种小分子敏感地感应和响应，从而有利于细菌的生存。通过结合CRISPR-Cas12a对单链DNA的切割能力和ATF对小分子和双链DNA的竞争结合活性，我们开发了一个简单、超灵敏、快速和高通量的小分子检测平台，称为CAT-SMelor(CRISPR-Cas12a和Tf介导型小分子检测器)。通过检测各种小分子的纳摩尔水平，包括尿酸和对羟基苯甲酸等结构相似的类似物，成功地评估了CAT-SMelor。我们还证明了我们的CAT-SMelor可以直接测量临床人体血液样本中的尿酸浓度，这表明CAT-SMelor在检测小分子方面具有很大的潜力。
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- andaTF-mediatedsmallmolecule detector). CaT-SMelor is successfully evaluated by detecting nanomolar levels of various small molecules, including uric acid andp-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.