期刊
ANALYTICA CHIMICA ACTA
卷 1125, 期 -, 页码 162-168出版社
ELSEVIER
DOI: 10.1016/j.aca.2020.05.017
关键词
CRISPR-Cas12a; DNA logic Gates; Biomolecular computing; Pathogenic bacteria sensing; Staphylococcus aureus
资金
- National Natural Science Foundation of China [81503086, 21672161]
- Tianjin Municipal Science and Technology Committee [18PTSYJC00140, 19JCYBJC27800]
For the first time, three 2-input elementary AND, OR, INHIBIT logic gates have been constructed by using CRISPR-Cas12a system. These logic gates utilised the intrinsic advantages of programmability, sequence specificity and high base resolution of CRISPR-Cas12a system. Among them, the AND gate owned the potentials as a built-in biosensor that responded rapidly to external pathogenic bacteria such as Staphylococcus aureus with high sensitivity and specificity. We applied the CRISPR-Cas12a based bacterial detection after a target-amplification using PCR. The total sample-to-answer time was appropriately 2.0 h, the limit of detection (LOD) was 10(3) CFU/mL, and the dynamic range was 10(3)-10(7) CFU/mL. Also, the sequence addressability enabled this AND logic gate to accurately trace back and distinguish input genes. These above-mentioned features were highly ideal to incur a rapid response to pathogenic bacteria for decision making. Our results not only validated the possibility of using CRISPR-Cas systems for constructing bio-computing devices but also provided a prototype of biosensor for rapid and intelligent pathogenic bacteria detection. (C) 2020 Elsevier B.V. All rights reserved.
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