A highly sensitive fluorometric method is described for the determination of microRNA-141. It is based on the use of arched probe-mediated isothermal exponential amplification reaction (EXPAR) and of DNA-templated silver nanoclusters (DNA-AgNCs). The EXPAR utilizes microRNA-141 as the trigger, polymerases and endonucleases as amplification activators, and two arched probes as exponential amplification templates. This enables the conversion of microRNA to a large number of reporter sequences under isothermal conditions within minutes. The generated reporter sequences act as scaffolds for the synthesis of fluorescent DNA-AgNCs by reduction of Ag (I) with NaBH4. The DNA-AgNCs function as signalling fluorophores with excitation/emission maxima at 540/610 nm. The method exhibits high sensitivity for microRNA-141 with a detection limit as low as 0.87 fM and a dynamic range from 1 fM to 500 fM. The method can distinguish nucleotides in the microRNA-200 family.
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Lee Y, Ahn C, Han J, Choi H, Kim J, Yim J, Lee J, Provost P, Radmark O, Kim S, Kim VN (2003) The nuclear RNase III Drosha initiates microRNA processing. Nature 425:415–419
Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297
Feng J, Xing W, Xie L (2016) Regulatory Roles of MicroRNAs in Diabetes. Int J Mol Sci 17:1729
Karnati HK, Panigrahi MK, Gutti RK, Greig NH, Tamargo IA (2015) miRNAs: Key Players in Neurodegenerative Disorders and Epilepsy. J Alzheimers Dis 48:563–580
Di Leva G, Garofalo M, Croce CM (2014) MicroRNAs in cancer. Annu Rev Pathol 9:287–314
Streit S, Michalski CW, Erkan M, Kleeff J, Friess H (2009) Northern blot analysis for detection and quantification of RNA in pancreatic cancer cells and tissues. Nat Protoc 4:37–43
Liu CG, Calin GA, Volinia S, Croce CM (2008) MicroRNA expression profiling using microarrays. Nat Protoc 3:563–578
Schmittgen TD, Lee EJ, Jiang J, Sarkar A, Yang L, Elton TS, Chen C (2008) Real-time PCR quantification of precursor and mature microRNA. Methods 44:31–38
Bi S, Yue S, Zhang S (2017) Hybridization chain reaction: a versatile molecular tool for biosensing, bioimaging, and biomedicine. Chem Soc Rev 46:4281–4298
Reid MS, Le XC, Zhang H (2018) Exponential Isothermal Amplification of Nucleic Acids and Assays for Proteins, Cells, Small Molecules, and Enzyme Activities: An EXPAR Example. Angew Chem Int Ed Engl 57:11856–11866
Dai W, Zhang J, Meng X, He J, Zhang K, Cao Y, Wang D, Dong H, Zhang X (2018) Catalytic hairpin assembly gel assay for multiple and sensitive microRNA detection. Theranostics 8:2646–2656
Li M, Xu X, Cai Q, Luo X, Zhou Z, Xu G, Xie Y (2019) Graphene oxide-based fluorometric determination of microRNA-141 using rolling circle amplification and exonuclease III-aided recycling amplification. Microchim Acta 186:531
Yu Y, Chen Z, Shi L, Yang F, Pan J, Zhang B, Sun D (2014) Ultrasensitive electrochemical detection of microRNA based on an arched probe mediated isothermal exponential amplification. Anal Chem 86:8200–8205
Liu YQ, Zhang M, Yin BC, Ye BC (2012) Attomolar ultrasensitive microRNA detection by DNA-scaffolded silver-nanocluster probe based on isothermal amplification. Anal Chem 84:5165–5169
Diez I, Ras RH (2011) Fluorescent silver nanoclusters. Nanoscale 3:1963–1970
Obliosca JM, Liu C, Yeh HC (2013) Fluorescent silver nanoclusters as DNA probes. Nanoscale 5:8443–8461
Berlina AN, Zherdev AV, Dzantiev BB (2019) Progress in rapid optical assays for heavy metal ions based on the use of nanoparticles and receptor molecules. Microchim Acta 186:172
Yuan Z, Chen YC, Li HW, Chang HT (2014) Fluorescent silver nanoclusters stabilized by DNA scaffolds. Chem Commun (Camb) 50:9800–9815
New SY, Lee ST, Su XD (2016) DNA-templated silver nanoclusters: structural correlation and fluorescence modulation. Nanoscale 8:17729–17746
Chen Y, Phipps ML, Werner JH, Chakraborty S, Martinez JS (2018) DNA Templated Metal Nanoclusters: From Emergent Properties to Unique Applications. Acc Chem Res 51:2756–2763
Mu WY, Yang R, Robertson A, Chen QY (2018) A near-infrared BSA coated DNA-AgNCs for cellular imaging. Colloids Surf B Biointerfaces 162:427–431
Shamsipur M, Pashabadi A, Molaabasi F, Hosseinkhani S (2017) Impedimetric monitoring of apoptosis using cytochrome-aptamer bioconjugated silver nanocluster. Biosens Bioelectron 90:195–202
Wu J, Li N, Yao Y, Tang D, Yang D, Ong'achwa Machuki J, Li J, Yu Y, Gao F (2018) DNA-Stabilized Silver Nanoclusters for Label-Free Fluorescence Imaging of Cell Surface Glycans and Fluorescence Guided Photothermal Therapy. Anal Chem 90:14368–14375
Zhang Y, Zhu C, Zhang L, Tan C, Yang J, Chen B, Wang L, Zhang H (2015) DNA-templated silver nanoclusters for multiplexed fluorescent DNA detection. Small 11:1385–1389
Borghei YS, Hosseini M, Ganjali MR, Ju H (2018) Colorimetric and energy transfer based fluorometric turn-on method for determination of microRNA using silver nanoclusters and gold nanoparticles. Microchim Acta 185:286
Jie G, Tan L, Zhao Y, Wang X (2017) A novel silver nanocluster in situ synthesized as versatile probe for electrochemiluminescence and electrochemical detection of thrombin by multiple signal amplification strategy. Biosens Bioelectron 94:243–249
Zhang Z, Guo C, Zhang S, He L, Wang M, Peng D, Tian J, Fang S (2017) Carbon-based nanocomposites with aptamer-templated silver nanoclusters for the highly sensitive and selective detection of platelet-derived growth factor. Biosens Bioelectron 89:735–742
Li Z, Ma YY, Wang J, Zeng XF, Li R, Kang W, Hao XK (2016) Exosomal microRNA-141 is upregulated in the serum of prostate cancer patients. Onco Targets Ther 9:139–148
Li R-D, Yin B-C, Ye B-C (2016) Ultrasensitive, colorimetric detection of microRNAs based on isothermal exponential amplification reaction-assisted gold nanoparticle amplification. Biosens Bioelectron 86:1011–1016
Tang Y, Liu M, Zhao Z, Li Q, Liang X, Tian J, Zhao S (2019) Fluorometric determination of microRNA-122 by using ExoIII-aided recycling amplification and polythymine induced formation of copper nanoparticles. Microchim Acta 186:133
Chen D, Wen S, Peng R, Gong Q, Fei J, Fu Z, Weng C, Liu M (2019) A triple signal amplification method for chemiluminescent detection of the cancer marker microRNA-21. Microchim Acta 186:410
Han Y, Qiu Z, Nawale GN, Varghese OP, Hilborn J, Tian B, Leifer K (2019) MicroRNA detection based on duplex-specific nuclease-assisted target recycling and gold nanoparticle/graphene oxide nanocomposite-mediated electrocatalytic amplification. Biosens Bioelectron 127:188–193
Wang J, Zhang L, Lu L, Kang T (2019) Molecular beacon immobilized on graphene oxide for enzyme-free signal amplification in electrochemiluminescent determination of microRNA. Microchim Acta 186:142
This work was supported by the Natural Science Foundation of Jiangsu Province (Grant No. BK20171144 and BK20161139) and the Scientific Research Foundation of Jiangsu Provincial Commission of Health and Family Planning, China (Grant No. H2018068).
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Wu, H., Wu, J., Liu, Y. et al. Fluorometric determination of microRNA using arched probe-mediated isothermal exponential amplification combined with DNA-templated silver nanoclusters. Microchim Acta 186, 715 (2019). https://doi.org/10.1007/s00604-019-3836-4
- Arched probe
- Silver nanoclusters
- Isothermal amplification
- DNA polymerase
- Nicking endonuclease
- Human serum