Citation: Jin Huang, Yong He, Xiao-Hai Yang, Ke Quan, Ke-Min Wang. Inhibited aptazyme-based catalytic molecular beacon for amplifi ed detection of adenosine[J]. Chinese Chemical Letters, ;2014, 25(9): 1211-1214. doi: 10.1016/j.cclet.2014.05.039 shu

Inhibited aptazyme-based catalytic molecular beacon for amplifi ed detection of adenosine

1.
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, China

Corresponding author: Ke-Min Wang,
Received Date: 12 March 2014
Available Online: 29 April 2014
Fund Project: Hunan Provincial Natural Science Foundation of China (No. 13JJ4032) (No. 2013M531779)the Fundamental Research Funds for the Central Universities of China. (No. 13JJ4032)

Combining the inhibited aptazyme and molecular beacon (MB), we developed a versatile sensing strategy for amplified detection of adenosine. In this strategy, the adenosine aptamer links to the 8-17 DNAzyme to form an aptazyme. A short sequence, denoted as inhibitor, is designed to form a duplex spanning the aptamer-DNAzyme junction, which blocks the catalytic function of the DNAzyme. Only in the presence of target adenosine, the aptamer binds to adenosine, thus the inhibitor dissociates from the aptamer portion of the aptazyme and can no longer form the stable duplex required to inhibit the catalytic activity of the aptazyme. The released DNAzyme domain will hybridize to the MB and catalyze the cleavage in the presence of Zn²⁺, making the fluorophore separate from the quencher and resulting in fluorescence signal. The results showed that the detection method has a dynamic range from 10 nmol/L to 1 nmol/L, with a detection limit of 10 nmol/L.
- Aptazyme,
- DNAzyme,
- Catalytic molecular beacon,
- Adenosine
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