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

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A CRISPR-Cas12a-derived biosensing platform for the highly sensitive detection of diverse small molecules.
Nature Communications ( IF 14.7 ) Pub Date : 2019-08-14 , DOI: 10.1038/s41467-019-11648-1
Mindong Liang _{1,

2} , Zilong Li ₃ , Weishan Wang ₃ , Jiakun Liu _{1,

4} , Leshi Liu ₁ , Guoliang Zhu ₁ , Loganathan Karthik ₁ , Man Wang ₅ , Ke-Feng Wang ₁ , Zhong Wang ₆ , Jing Yu ₇ , Yuting Shuai ₁ , Jiaming Yu ₁ , Lu Zhang ₁ , Zhiheng Yang ₁ , Chuan Li ₁ , Qian Zhang ₁ , Tong Shi ₁ , Liming Zhou ₁ , Feng Xie ₃ , Huanqin Dai ₃ , Xueting Liu ₁ , Jingyu Zhang ₁ , Guang Liu ₁ , Ying Zhuo ₁ , Buchang Zhang ₂ , Chenli Liu ₄ , Shanshan Li ₈ , Xuekui Xia ₉ , Yaojun Tong ₁₀ , Yanwen Liu ₁₁ , Gil Alterovitz ₁₂ , Gao-Yi Tan ₁ , Li-Xin Zhang ₁

Affiliation

State Key Laboratory of Bioreactor Engineering, and School of Biotechnology, East China University of Science and Technology (ECUST), 200237, Shanghai, China.
Institute of Physical Science and Information Technology, Anhui University, 230601, Hefei, China.
State Key Laboratory of Microbial Resources and CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China.
Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, 518055, Shenzhen, China.
School of Agriculture and Biology, Shanghai Jiao Tong University, 200240, Shanghai, China.
Department of Urology, Sixth People's Hospital South Campus Affiliated to Shanghai Jiao Tong University, 201499, Shanghai, China.
Department of Clinical Laboratory, Sixth People's Hospital South Campus Affiliated to Shanghai Jiao Tong University, Shanghai, 201499, China.
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, China.
Key Biosensor Laboratory of Shandong Province, Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), 250013, Jinan, China.
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs, Lyngby, Denmark.
inBiome Sciences, 3403 SW 86th ST, Gainesville, FL, 32608, USA.
Harvard Medical School Countway Library, 10 Shattuck Street, Boston, MA, 02115, USA.

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.

中文翻译：

CRISPR-Cas12a衍生的生物传感平台，可高度灵敏地检测各种小分子。

除了基因组编辑外，CRISPR-Cas12a最近还被用于具有attomolar敏感性的DNA检测应用，但据我们所知，它尚未被用于检测小分子。细菌变构转录因子（aTFs）已经进化为可以感知并敏感地响应各种小分子，从而有利于细菌存活。通过结合CRISPR-Cas12a的单链DNA切割能力和aTFs对小分子和双链DNA的竞争结合活性，我们在此开发了一种简单，超灵敏，快速且高通量的平台，用于检测小分子，称为CaT-SMelor（CRISPR-Cas12a和aTF介导的小分子检测器）。通过检测各种小分子的纳摩尔水平，成功评估了CaT-SMelor，在结构上类似的类似物中包括尿酸和对羟基苯甲酸。我们还证明，我们的CaT-SMelor直接测量临床人血样品中的尿酸浓度，表明CaT-SMelor在检测小分子方面具有巨大潜力。

更新日期：2019-08-14

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