A Graphene-enhanced imaging of microRNA with enzyme-free signal amplification of catalyzed hairpin assembly in living cells.

In situ imaging of miRNA in living cells could help us to monitor the miRNA expression in real time and obtain accurate information for studying miRNA related bioprocesses and disease. Given the low-level expression of miRNA, amplification strategies for intracellular miRNA are imperative. Here, we propose an amplification strategy with a non-destructive enzyme-free manner in living cells using catalyzed hairpin assembly (CHA) based on graphene oxide (GO) for cellular miRNA imaging. The enzyme-free CHA exhibits stringent recognition and excellent signal amplification of miRNA in the living cells. GO is a good candidate as a fluorescence quencher and cellular carrier. Taking the advantages of the CHA and GO, we can monitor the miRNA at low level in living cells with a simple, sensitive and real-time manner. Finally, imaging of miRNAs in the different expression cells is realized. The novel method could supply an effective tool to visualize intracellular low-level miRNAs and help us to further understand the role of miRNAs in cellular processes.

[1]  Longhua Tang,et al.  Duplex DNA/Graphene Oxide Biointerface: From Fundamental Understanding to Specific Enzymatic Effects , 2012 .

[2]  Debin Zhu,et al.  Detection of microRNA in clinical tumor samples by isothermal enzyme-free amplification and label-free graphene oxide-based SYBR Green I fluorescence platform. , 2015, Biosensors & bioelectronics.

[3]  Adam Grundhoff,et al.  A combined computational and microarray-based approach identifies novel microRNAs encoded by human gamma-herpesviruses. , 2006, RNA.

[4]  Qian Wang,et al.  High specific and ultrasensitive isothermal detection of microRNA by padlock probe-based exponential rolling circle amplification. , 2013, Analytical chemistry.

[5]  G. Seelig,et al.  Enzyme-Free Nucleic Acid Logic Circuits , 2022 .

[6]  Ying Wang,et al.  The graphene/nucleic acid nanobiointerface. , 2015, Chemical Society reviews.

[7]  Kwang S. Kim,et al.  Noncovalent Functionalization of Graphene and Graphene Oxide for Energy Materials, Biosensing, Catalytic, and Biomedical Applications. , 2016, Chemical reviews.

[8]  Qian Wang,et al.  Graphene fluorescence switch-based cooperative amplification: a sensitive and accurate method to detection microRNA. , 2014, Analytical chemistry.

[9]  Yun Chen,et al.  Quantification of microRNA by DNA-Peptide Probe and Liquid Chromatography-Tandem Mass Spectrometry-Based Quasi-Targeted Proteomics. , 2016, Analytical chemistry.

[10]  C. Kang,et al.  AC1MMYR2 impairs high dose paclitaxel-induced tumor metastasis by targeting miR-21/CDK5 axis. , 2015, Cancer letters.

[11]  Yu-Qiang Liu,et al.  One-step, multiplexed fluorescence detection of microRNAs based on duplex-specific nuclease signal amplification. , 2012, Journal of the American Chemical Society.

[12]  James W Jacobson,et al.  MicroRNA: Potential for Cancer Detection, Diagnosis, and Prognosis. , 2007, Cancer research.

[13]  Brian S. Roberts,et al.  Simple, quantitative primer-extension PCR assay for direct monitoring of microRNAs and short-interfering RNAs. , 2005, RNA.

[14]  Zhigang Li,et al.  Enzyme-free and amplified fluorescence DNA detection using bimolecular beacons. , 2012, Analytical chemistry.

[15]  Yuehe Lin,et al.  Aptamer/graphene oxide nanocomplex for in situ molecular probing in living cells. , 2010, Journal of the American Chemical Society.

[16]  C. Harris,et al.  Genetic variation in microRNA networks: the implications for cancer research , 2010, Nature Reviews Cancer.

[17]  Francois Natt,et al.  A novel microarray approach reveals new tissue-specific signatures of known and predicted mammalian microRNAs , 2007, Nucleic acids research.

[18]  John J Rossi,et al.  New Hope for a MicroRNA Therapy for Liver Cancer , 2009, Cell.

[19]  É. Várallyay,et al.  Detection of microRNAs by Northern blot analyses using LNA probes. , 2007, Methods.

[20]  Lei Han,et al.  AC1MMYR2, an inhibitor of dicer-mediated biogenesis of Oncomir miR-21, reverses epithelial-mesenchymal transition and suppresses tumor growth and progression. , 2013, Cancer research.

[21]  Won Jun Kang,et al.  Dual optical biosensors for imaging microRNA-1 during myogenesis. , 2012, Biomaterials.

[22]  Yu Chen,et al.  Two-dimensional graphene analogues for biomedical applications. , 2015, Chemical Society reviews.

[23]  Longhua Tang,et al.  DNA-directed self-assembly of graphene oxide with applications to ultrasensitive oligonucleotide assay. , 2011, ACS nano.

[24]  Jingmin Jin,et al.  Rapid electronic detection of probe-specific microRNAs using thin nanopore sensors. , 2010, Nature nanotechnology.

[25]  L. Druhan,et al.  Forced Expression of Heat Shock Protein 27 (Hsp27) Reverses P-Glycoprotein (ABCB1)-mediated Drug Efflux and MDR1 Gene Expression in Adriamycin-resistant Human Breast Cancer Cells* , 2011, The Journal of Biological Chemistry.

[26]  Rui Shi,et al.  Facile means for quantifying microRNA expression by real-time PCR. , 2005, BioTechniques.

[27]  Feng Yan,et al.  The use of polyethylenimine-grafted graphene nanoribbon for cellular delivery of locked nucleic acid modified molecular beacon for recognition of microRNA. , 2011, Biomaterials.

[28]  Cuichen Wu,et al.  A Nonenzymatic Hairpin DNA Cascade Reaction Provides High Signal Gain of mRNA Imaging inside Live Cells , 2015, Journal of the American Chemical Society.

[29]  Hongwei Gong,et al.  Programmable Mg(2+)-dependent DNAzyme switch by the catalytic hairpin DNA assembly for dual-signal amplification toward homogeneous analysis of protein and DNA. , 2015, Chemical communications.

[30]  Yuehe Lin,et al.  In situ simultaneous monitoring of ATP and GTP using a graphene oxide nanosheet–based sensing platform in living cells , 2014, Nature Protocols.

[31]  Y. Mo,et al.  Negative regulation of lncRNA GAS5 by miR-21 , 2013, Cell Death and Differentiation.

[32]  S. Lowe,et al.  A microRNA polycistron as a potential human oncogene , 2005, Nature.

[33]  Yongqiang Cheng,et al.  Ultrasensitive detection of microRNAs by exponential isothermal amplification. , 2010, Angewandte Chemie.

[34]  Jinghong Li,et al.  Sensitive and rapid screening of T4 polynucleotide kinase activity and inhibition based on coupled exonuclease reaction and graphene oxide platform. , 2011, Analytical chemistry.

[35]  C. Sawyers The cancer biomarker problem , 2008, Nature.

[36]  Huang-Hao Yang,et al.  Increasing the sensitivity and single-base mismatch selectivity of the molecular beacon using graphene oxide as the "nanoquencher". , 2010, Chemistry.

[37]  Longhua Tang,et al.  Toehold-initiated rolling circle amplification for visualizing individual microRNAs in situ in single cells. , 2014, Angewandte Chemie.

[38]  Sang Woo Han,et al.  Quantitative and multiplexed microRNA sensing in living cells based on peptide nucleic acid and nano graphene oxide (PANGO). , 2013, ACS nano.

[39]  K. Livak,et al.  Real-time quantification of microRNAs by stem–loop RT–PCR , 2005, Nucleic acids research.

[40]  C. Croce,et al.  An oligonucleotide microchip for genome-wide microRNA profiling in human and mouse tissues. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[41]  Nóra Varga,et al.  Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes. , 2004, Nucleic acids research.

[42]  Kai Yang,et al.  Nano-graphene in biomedicine: theranostic applications. , 2013, Chemical Society reviews.

[43]  F. Slack,et al.  RAS Is Regulated by the let-7 MicroRNA Family , 2005, Cell.

[44]  H. Ju,et al.  Fluorescence resonance energy transfer between quantum dots and graphene oxide for sensing biomolecules. , 2010, Analytical chemistry.

[45]  Chuan He,et al.  Live Cell MicroRNA Imaging Using Cascade Hybridization Reaction. , 2015, Journal of the American Chemical Society.

[46]  Da Chen,et al.  Graphene oxide: preparation, functionalization, and electrochemical applications. , 2012, Chemical reviews.

[47]  C. Croce,et al.  MicroRNA signatures in human cancers , 2006, Nature Reviews Cancer.

[48]  Andrea Ventura,et al.  MicroRNAs and Cancer: Short RNAs Go a Long Way , 2009, Cell.

[49]  Soonhag Kim,et al.  Molecular imaging of a cancer-targeting theragnostics probe using a nucleolin aptamer- and microRNA-221 molecular beacon-conjugated nanoparticle. , 2012, Biomaterials.