A nuclease-assisted label-free aptasensor for fluorescence turn-on detection of ATP based on the in situ formation of copper nanoparticles.

Owing to their promising advantages in biochemical analysis, aptamer-based sensing systems for the fluorescence detection of important biomolecules are being extensively investigated. Herein, we propose a turn-on fluorescent aptasensor for label-free detection of adenosine triphosphate (ATP) by utilizing the in situ formation of copper nanoparticles (CuNPs) and the specific digestion capability of exonuclease I (Exo I). In this assay, the addition of ATP can effectively hinder the digestion of aptamer-derived oligonucleotides due to the G-quadruplex structure. Accordingly, the remaining poly thymine at 5'-terminus of substrate DNA can serve as an efficient template for red-emitting fluorescent CuNPs with a Mega-Stokes shifting in buffered solution, which can be used to evaluate the concentration of ATP. This method is cost-effective and facile, because it avoids the use of traditional dye-labeled DNA strands and complex operation steps. Under optimized conditions, this method achieves a selective response for ATP with a detection limit of 93nM, and exhibits a good detection performance in biological samples.

[1]  Andriy Mokhir,et al.  Selective dsDNA-templated formation of copper nanoparticles in solution. , 2010, Angewandte Chemie.

[2]  Yongming Guo,et al.  Fluorescent copper nanoparticles: recent advances in synthesis and applications for sensing metal ions. , 2016, Nanoscale.

[3]  Kemin Wang,et al.  Poly(thymine)-templated selective formation of fluorescent copper nanoparticles. , 2013, Angewandte Chemie.

[4]  Chun-yang Zhang,et al.  Real-time monitoring of small biological molecules by ligation-mediated polymerase chain reaction. , 2015, Chemical communications.

[5]  Hai-Bo Wang,et al.  A fluorescent biosensor for protein detection based on poly(thymine)-templated copper nanoparticles and terminal protection of small molecule-linked DNA. , 2015, Biosensors & bioelectronics.

[6]  Michael Famulok,et al.  Aptamer modules as sensors and detectors. , 2011, Accounts of chemical research.

[7]  Bingling Li,et al.  "Fitting" makes "sensing" simple: label-free detection strategies based on nucleic acid aptamers. , 2013, Accounts of chemical research.

[8]  Meng Zhao,et al.  A label-free aptasensor for highly sensitive detection of ATP and thrombin based on metal-enhanced PicoGreen fluorescence. , 2015, Biosensors & bioelectronics.

[9]  Z. Li,et al.  A label-free method for detecting biothiols based on poly(thymine)-templated copper nanoparticles. , 2015, Biosensors & bioelectronics.

[10]  Shenghao Xu,et al.  Sequence-dependent dsDNA-templated formation of fluorescent copper nanoparticles. , 2015, Chemistry.

[11]  Louai Labanieh,et al.  Nucleic acid aptamers in cancer research, diagnosis and therapy. , 2015, Chemical Society reviews.

[12]  Xi Chen,et al.  DNA-based ATP sensing , 2016 .

[13]  J. Szostak,et al.  A DNA aptamer that binds adenosine and ATP. , 1995, Biochemistry.

[14]  D. He,et al.  A fluorescent aptasensor for amplified label-free detection of adenosine triphosphate based on core-shell Ag@SiO2 nanoparticles. , 2016, Biosensors & bioelectronics.

[15]  Chen Su,et al.  Enzymatic polymerization of poly(thymine) for the synthesis of copper nanoparticles with tunable size and their application in enzyme sensing. , 2015, Chemical communications.

[16]  Jin-Ook Baeg,et al.  Label-free dual assay of DNA sequences and potassium ions using an aptamer probe and a molecular light switch complex. , 2009, Chemical communications.

[17]  Kemin Wang,et al.  Poly(thymine)-templated fluorescent copper nanoparticles for ultrasensitive label-free nuclease assay and its inhibitors screening. , 2013, Analytical chemistry.

[18]  Y. Jong,et al.  Label-Free Fluorescent Copper Nanoclusters for Genotyping of Deletion and Duplication of Duchenne Muscular Dystrophy. , 2015, Analytical chemistry.

[19]  Xiaojun Zhang,et al.  A label-free and enzyme-free ultra-sensitive transcription factors biosensor using DNA-templated copper nanoparticles as fluorescent indicator and hairpin DNA cascade reaction as signal amplifier. , 2016, Biosensors & bioelectronics.

[20]  Audrey Sassolas,et al.  Optical detection systems using immobilized aptamers. , 2011, Biosensors & bioelectronics.

[21]  Kemin Wang,et al.  dsDNA-specific fluorescent copper nanoparticles as a "green" nano-dye for polymerization-mediated biochemical analysis. , 2014, Chemical communications.

[22]  P. Tinnefeld,et al.  Multifunctional Dumbbell-Shaped DNA-Templated Selective Formation of Fluorescent Silver Nanoclusters or Copper Nanoparticles for Sensitive Detection of Biomolecules. , 2016, ACS applied materials & interfaces.

[23]  Shachar Richter,et al.  Bio-inspired synthesis of chiral silver nanoparticles in mucin glycoprotein--the natural choice. , 2011, Chemical communications.

[24]  Chuan Dong,et al.  An exonuclease I-based label-free fluorometric aptasensor for adenosine triphosphate (ATP) detection with a wide concentration range. , 2015, Biosensors & bioelectronics.

[25]  Meiping Zhao,et al.  In-vivo fluorescence imaging of adenosine 5′-triphosphate , 2016 .

[26]  S. Dong,et al.  Double-strand DNA-templated formation of copper nanoparticles as fluorescent probe for label-free aptamer sensor. , 2011, Analytical chemistry.