Cross-catalytic hairpin assembly-based exponential signal amplification for CRET assay with low background noise
| Autor: | Tingting Zhao, Sai Bi, Yongcun Yan, Shuzhen Yue, Hongjie Qi |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Luminescence
Kinetics Biomedical Engineering Biophysics Nanotechnology Biosensing Techniques 02 engineering and technology 010402 general chemistry 01 natural sciences Signal Catalysis law.invention chemistry.chemical_compound Exponential growth law DNA nanotechnology Electrochemistry Chemiluminescence Inverted Repeat Sequences DNA General Medicine 021001 nanoscience & nanotechnology 0104 chemical sciences MicroRNAs chemistry Luminescent Measurements Nucleic acid Nucleic Acid Conformation 0210 nano-technology Biosensor Biotechnology |
| Zdroj: | Biosensors and Bioelectronics. 94:671-676 |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/j.bios.2017.03.071 |
| Popis: | A toehold-mediated strand displacement (TMSD)-based cross-catalytic hairpin assembly (C-CHA) is demonstrated in this study, achieving exponential amplification of nucleic acids. Functionally, this system consists of four hairpins (H1, H2, H3 and H4) and one single-stranded initiator (I). Upon the introduction of I, the first CHA reaction (CHA1) is triggered, leading to the self-assembly of hybrid H1·H2 that then initiates the second CHA reaction (CHA2) to obtain the hybrid H3·H4. Since the single-stranded region in H3·H4 is identical to I, a new CHA1 is initiated, which thus achieves cross operation of CHA1 and CHA2 and exponential growth kinetics. Interestingly, because the C-CHA performs in a cascade manner, this system can be considered as multi-level molecular logic circuits with feedback mechanism. Moreover, through incorporating G-quadruplex subunits and fluorescein isothiocyanate (FITC) in the product of H1·H2, this C-CHA is readily utilized to fabricate a chemiluminescence resonance energy transfer (CRET) biosensing platform, achieving sensitive and selective detection of DNA and microRNA in real samples. Since the high background signal induced by FITC in the absence of initiator is greatly reduced through labeling quencher in H1, the signal-to-noise ratio and detection sensitivity are improved significantly. Therefore, our proposed C-CHA protocol holds a great potential for further applications in not only building complex autonomous systems but also the development of biosensing platforms and DNA nanotechnology. |
| Databáze: | OpenAIRE |
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