In Situ Measurement of ATP in Single Cells by an Amphiphilic Aptamer-Assisted Electrochemical Nano-Biosensor.

In situ and quantitative measurements of adenosine 5'-triphosphate (ATP) in single living cells are highly desired for understanding several sorts of necessary physiological and pathological processes. Due to its small size and high sensitivity, an ultra-microelectrode can be used for single-cell analysis. However, ATP is difficult to detect in single cells because it is nonelectroactive and low in content. Herein, we introduced an electrochemical nano-biosensor based on an amphiphilic aptamer-assisted carbon fiber nanoelectrode (aptCFNE) with high signal-to-noise ratio. The low current (e.g., 60 pA) and the tiny diameter of the tip (ca. 400 nm) of the nanosensor made it noninvasive to living cells. The amphiphilic aptamer has good biocompatibility and can be stably modified to the surface of functionalized electrodes. CFNE, which was modified with ferrocene-labeled aptamer, could quickly and selectively detect ATP content in the nucleus, cytoplasm, and extracellular space of single HeLa cells. The results showed that the ATP contents in the nucleus, cytoplasm, and extracellular space were 568 ± 9, 461 ± 20, and 312 ± 4 μM, respectively. The anticancer drug treatment effects on the cellular level were further recorded, which was of great significance for understanding ATP-related biological processes and drug screenings. This strategy is universally applicable to detect other targets by changing the aptamer sequence, which will greatly improve our understanding of cell heterogeneity and provide a more reliable scientific basis for exploring major diseases at the single-cell level.

[1]  Xiangzhi Cui,et al.  Modulation of mitochondrial electron transport chain by pyroptosis nanoagonists for photoresponsive tumor destruction , 2022, Nano Today.

[2]  U. Häfeli,et al.  Aptamers used for molecular imaging and theranostics - recent developments , 2022, Theranostics.

[3]  Zachary D. Schultz,et al.  High-Throughput Surface-Enhanced Raman Scattering for Screening Chemical Sensor Candidates Enabled by Bipolar Electrochemistry. , 2022, ACS sensors.

[4]  A. Ewing,et al.  Single‐Vesicle Electrochemistry Following Repetitive Stimulation Reveals a Mechanism for Plasticity Changes with Iron Deficiency , 2022, Angewandte Chemie.

[5]  A. Escarpa,et al.  New trends in enzyme-free electrochemical sensing of ROS/RNS. Application to live cell analysis , 2022, Microchimica Acta.

[6]  A. Ewing,et al.  Simultaneous Counting of Molecules in the Halo and Dense‐Core of Nanovesicles by Regulating Dynamics of Vesicle Opening , 2022, Angewandte Chemie.

[7]  Liangqia Guo,et al.  Salt-template preparation of Mo5N6 nanosheets with peroxidase- and catalase-like activities and application for colorimetric determination of 4-aminophenol , 2021, Microchimica Acta.

[8]  Zhaoming Li,et al.  Synergistic anticancer effect of a combination of chidamide and etoposide against NK/T cell lymphoma , 2021, Hematological oncology.

[9]  Xiaoding Xu,et al.  A genetically encoded fluorescent biosensor for monitoring ATP in living cells with heterobifunctional aptamers. , 2021, Biosensors & bioelectronics.

[10]  M. Tokeshi,et al.  Electrochemical enzyme-based blood ATP and lactate sensor for a rapid and straightforward evaluation of illness severity , 2021, Biosensors and Bioelectronics.

[11]  Hongjie Xiong,et al.  Engineered Aptamer-Organic Amphiphile Self-Assemblies for Biomedical Applications: Progress and Challenges. , 2021, Small.

[12]  Fuan Wang,et al.  A Cooperatively Activatable DNA Nanoprobe for Cancer Cell-Selective Imaging of ATP. , 2021, Analytical chemistry.

[13]  Yan Zeng,et al.  Discriminative intracellular and extracellular ATP detection based on magnetically controlled antimicrobial peptide , 2021 .

[14]  S. Marinesco.  Micro- and nanoelectrodes for neurotransmitter monitoring , 2021 .

[15]  Juewen Liu,et al.  Nanomaterial and Aptamer-Based Sensing: Target Binding versus Target Adsorption Illustrated by the Detection of Adenosine and ATP on Metal Oxides and Graphene Oxide. , 2021, Analytical chemistry.

[16]  Hongyuan Chen,et al.  An Integrated Electrochemical Nanodevice for Intracellular RNA Collection and Detection in Single Living Cell. , 2020, Angewandte Chemie.

[17]  Hong-Wu Tang,et al.  Detection of ATP from “fluorescence” to “enhanced fluorescence” based on metal-enhanced fluorescence triggered by aptamer nanoswitch , 2020 .

[18]  Jiajia Song,et al.  Electrochemical Nanoaptasensor for Continuous Monitoring ATP Fluctuation at Subcellular Level. , 2020, Analytical chemistry.

[19]  Jingbo Hu,et al.  A Generalizable and Noncovalent Strategy of Interfacing Aptamers with Microelectrode for Selective Sensing of Neurotransmitter In Vivo. , 2020, Angewandte Chemie.

[20]  Kecheng Zhang,et al.  Fluorescent carbon dots for in situ monitoring of lysosomal ATP levels. , 2020, Analytical chemistry.

[21]  Wenjie Ma,et al.  Unveiling the Role of DJ-1 Protein in Vesicular Storage and Release of Catecholamine with Nano/Micro-Tip Electrodes. , 2020, Angewandte Chemie.

[22]  T. Scheibel,et al.  Functionalized DNA-spider silk nanohydrogels for controlled protein binding and release , 2020, Materials today. Bio.

[23]  L. Mao,et al.  Enzymatic electrochemical biosensors for in situ neurochemical measurement , 2020, Current Opinion in Electrochemistry.

[24]  F. Tezcan,et al.  Electron Transfer in Nitrogenase. , 2020, Chemical reviews.

[25]  Xiaojing Liu,et al.  A cell membrane-anchored DNA nano-platform for programming cellular interactions. , 2019, Journal of the American Chemical Society.

[26]  R. Tampé,et al.  ABCE1 Controls Ribosome Recycling by an Asymmetric Dynamic Conformational Equilibrium , 2019, Cell reports.

[27]  M. L. Yola,et al.  Enhanced surface plasmon resonance (SPR) signals based on immobilization of core-shell nanoparticles incorporated boron nitride nanosheets: Development of molecularly imprinted SPR nanosensor for anticancer drug, etoposide. , 2019, Biosensors & bioelectronics.

[28]  Yong Wang,et al.  Assembly of Bifunctional Aptamer-Fibrinogen Macromer for VEGF Delivery and Skin Wound Healing. , 2019, Chemistry of materials : a publication of the American Chemical Society.

[29]  Hitoshi Shiku,et al.  Intracellular Electrochemical Sensing , 2018, Electroanalysis.

[30]  M. Raghunath,et al.  RGB-Color Intensiometric Indicators to Visualize Spatiotemporal Dynamics of ATP in Single Cells. , 2018, Angewandte Chemie.

[31]  Ping Yu,et al.  Photoinduced Regeneration of an Aptamer-Based Electrochemical Sensor for Sensitively Detecting Adenosine Triphosphate. , 2018, Analytical chemistry.

[32]  R. Metzler,et al.  Manipulation and Motion of Organelles and Single Molecules in Living Cells. , 2017, Chemical reviews.

[33]  W. Koh,et al.  Single-cell genome sequencing: current state of the science , 2016, Nature Reviews Genetics.

[34]  Takeharu Nagai,et al.  Visualization of ATP levels inside single living cells with fluorescence resonance energy transfer-based genetically encoded indicators , 2009, Proceedings of the National Academy of Sciences.

[35]  E. Wang,et al.  A strategy for constructing a hybrid bilayer membrane based on a carbon substrate. , 2003, Analytical chemistry.

[36]  M. Porter,et al.  Electrochemical oxidation of amine-containing compounds. A route to the surface modification of glassy carbon electrodes , 1994 .