Abstract
MicroRNAs (miRNAs) are stably present in circulatory systems. They are bound to various carriers like proteins, lipoprotein particles, and exosomes. Investigating the process of miRNA distribution among these carriers will help improve our understanding of their functions in the extracellular environment and their potential relationship with diseases. Here, we describe how to obtain the distribution profiles of circulating miRNAs by separation of different miRNA carriers in human serum with asymmetrical flow field flow fractionation (AF4), and detection of the miRNAs in the eluted fractions that enrich particular types of carriers with RT-qPCR.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Hagen JW, Lai EC (2008) microRNA control of cell-cell signaling during development and disease. Cell Cycle 7:2327–2332
Nicoloso MS, Spizzo R, Shimizu M, Rossi S, Calin GA (2009) MicroRNAs—the micro steering wheel of tumour metastases. Nat Rev Cancer 9:293–302. doi:10.1038/nrc2619
Shi M, Liu D, Duan H, Shen B, Guo N (2010) Metastasis-related miRNAs, active players in breast cancer invasion, and metastasis. Cancer Metastasis Rev 29:785–799. doi:10.1007/s10555-010-9265-9
Volinia S, Galasso M, Costinean S, Tagliavini L, Gamberoni G, Drusco A, Marchesini J, Mascellani N, Sana ME, Abu Jarour R, Desponts C, Teitell M, Baffa R, Aqeilan R, Iorio MV, Taccioli C, Garzon R, Di Leva G, Fabbri M, Catozzi M, Previati M, Ambs S, Palumbo T, Garofalo M, Veronese A, Bottoni A, Gasparini P, Harris CC, Visone R, Pekarsky Y, de la Chapelle A, Bloomston M, Dillhoff M, Rassenti LZ, Kipps TJ, Huebner K, Pichiorri F, Lenze D, Cairo S, Buendia M-A, Pineau P, Dejean A, Zanesi N, Rossi S, Calin GA, Liu C-G, Palatini J, Negrini M, Vecchione A, Rosenberg A, Croce CM (2010) Reprogramming of miRNA networks in cancer and leukemia. Genome Res 20:589–599. doi:10.1101/gr.098046.109
Dontu G, de Rinaldis E (2010) MicroRNAs: shortcuts in dealing with molecular complexity? Breast Cancer Res 12:301. doi:10.1186/bcr2455
Ma L, Weinberg RA (2008) Micromanagers of malignancy: role of microRNAs in regulating metastasis. Trends Genet 24:448–456. doi:10.1016/j.tig.2008.06.004
Ventura A, Jacks T (2009) MicroRNAs and cancer: short RNAs go a long way. Cell 136:586–591. doi:10.1016/j.cell.2009.02.005
Creemers EE, Tijsen AJ, Pinto YM (2012) Circulating microRNAs: novel biomarkers and extracellular communicators in cardiovascular disease? Circ Res 110:483–495. doi:10.1161/CIRCRESAHA.111.247452
Laganà A, Russo F, Veneziano D, Bella SD, Giugno R, Pulvirenti A, Croce CM, Ferro A (2013) Extracellular circulating viral microRNAs: current knowledge and perspectives. Front Genet. doi:10.3389/fgene.2013.00120
Olivieri F, Rippo MR, Procopio AD, Fazioli F (2013) Circulating inflamma-miRs in aging and age-related diseases. Front Genet 4:121. doi:10.3389/fgene.2013.00121
Rykova EY, Laktionov PP, Vlassov VV (2010) Circulating nucleic acids in health and disease. In: Kikuchi Y, Rykova EY (eds) Extracellular nucleic acids. Springer, Berlin, Heidelberg, pp 93–128
Etheridge A, Lee I, Hood L, Galas D, Wang K (2011) Extracellular microRNA: a new source of biomarkers. Mutat Res 717:85–90. doi:10.1016/j.mrfmmm.2011.03.004
Turchinovich A, Weiz L, Burwinkel B (2012) Extracellular miRNAs: the mystery of their origin and function. Trends Biochem Sci 37:460–465. doi:10.1016/j.tibs.2012.08.003
Wang K, Zhang S, Weber J, Baxter D, Galas DJ (2010) Export of microRNAs and microRNA-protective protein by mammalian cells. Nucleic Acids Res gkq601. doi: 10.1093/nar/gkq601
Wahlund KG, Giddings JC (1987) Properties of an asymmetrical flow field-flow fractionation channel having one permeable wall. Anal Chem 59:1332–1339. doi:10.1021/ac00136a016
Kim KH, Moon MH (2011) Chip-type asymmetrical flow field-flow fractionation channel coupled with mass spectrometry for top-down protein identification. Anal Chem 83:8652–8658. doi:10.1021/ac202098b
Oh S, Kang D, Ahn S-M, Simpson RJ, Lee B-H, Moon MH (2007) Miniaturized asymmetrical flow field-flow fractionation: application to biological vesicles. J Sep Sci 30:1082–1087
Kim KH, Lee JY, Lim S, Moon MH (2013) Top-down lipidomic analysis of human lipoproteins by chip-type asymmetrical flow field-flow fractionation-electrospray ionization-tandem mass spectrometry. J Chromatogr A 1280:92–97. doi:10.1016/j.chroma.2013.01.025
Madörin M, van Hoogevest P, Hilfiker R, Langwost B, Kresbach GM, Ehrat M, Leuenberger H (1997) Analysis of drug/plasma protein interactions by means of asymmetrical flow field-flow fractionation. Pharm Res 14:1706–1712
Pollastrini J, Dillon TM, Bondarenko P, Chou RY-T (2011) Field flow fractionation for assessing neonatal Fc receptor and Fcγ receptor binding to monoclonal antibodies in solution. Anal Biochem 414:88–98. doi:10.1016/j.ab.2011.03.001
Schachermeyer S, Ashby J, Zhong W (2013) Aptamer-protein binding detected by asymmetric flow field flow fractionation. J Chromatogr A 1295:107–113. doi:10.1016/j.chroma.2013.04.063
Ashby J, Flack K, Jimenez LA, Duan Y, Khatib A-K, Somlo G, Wang SE, Cui X, Zhong W (2014) Distribution profiling of circulating microRNAs in serum. Anal Chem 86:9343–9349. doi:10.1021/ac5028929
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Flack, K., Jimenez, L.A., Zhong, W. (2017). Analysis of the Distribution Profiles of Circulating MicroRNAs by Asymmetrical Flow Field Flow Fractionation. In: Rani, S. (eds) MicroRNA Profiling. Methods in Molecular Biology, vol 1509. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6524-3_15
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6524-3_15
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6522-9
Online ISBN: 978-1-4939-6524-3
eBook Packages: Springer Protocols