Abstract
MicroRNAs (miRNAs) are a class of small, highly conserved, and noncoding RNAs that modulate gene expression by regulating the activity and stability of target mRNAs. MiRNAs play significant roles by controlling fundamental cellular processes and its deregulation is associated with various diseases. Ubiquitous expression and its release into circulation make them interesting biomarkers, which can be measured by different platforms. In this book chapter, we provide a specific protocol that describes the detection of circulating miRNAs in CSF by using RT-qPCR.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Weber JA, Baxter DH, Zhang S et al (2010) The microRNA spectrum in 12 body fluids. Clin Chem 56(11):1733–1741. https://doi.org/10.1373/clinchem.2010.147405
Mitchell PS, Parkin RK, Kroh EM et al (2008) Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A 105(30):10513–10518. https://doi.org/10.1073/pnas.0804549105
Balusu S, Van Wonterghem E, De Rycke R et al (2016) Identification of a novel mechanism of blood-brain communication during peripheral inflammation via choroid plexus-derived extracellular vesicles. EMBO Mol Med 8(10):1162–1183. https://doi.org/10.15252/emmm.201606271
Pritchard CC, Cheng HH, Tewari M (2012) MicroRNA profiling: approaches and considerations. Nat Rev Genet 13(5):358–369. https://doi.org/10.1038/nrg3198
Mestdagh P, Hartmann N, Baeriswyl L et al (2014) Evaluation of quantitative miRNA expression platforms in the microRNA quality control (miRQC) study. Nat Methods 11(8):809–815. https://doi.org/10.1038/nmeth.3014
Witwer KW, Halushka MK (2016) Toward the promise of microRNAs—enhancing reproducibility and rigor in microRNA research. RNA Biol 13(11):1103–1116. https://doi.org/10.1080/15476286.2016.1236172
Bustin SA, Benes V, Garson JA et al (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55(4):611–622. https://doi.org/10.1373/clinchem.2008.112797
Bustin SA (2010) Why the need for qPCR publication guidelines? The case for MIQE. Methods 50(4):217–226. https://doi.org/10.1016/j.ymeth.2009.12.006
Bustin SA, Beaulieu JF, Huggett J et al (2010) MIQE précis: practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments. BMC Mol Biol 11:74. https://doi.org/10.1186/1471-2199-11-74
Kirschner MB, van Zandwijk N, Reid G (2013) Cell-free microRNAs: potential biomarkers in need of standardized reporting. Front Genet 4:56. https://doi.org/10.3389/fgene.2013.00056
Kirschner MB, Edelman JJB, Kao SCH et al (2013) The impact of hemolysis on cell-free microRNA biomarkers. Front Genet 4:94. https://doi.org/10.3389/fgene.2013.00094, Article No.: 94
Mestdagh P, Van Vlierberghe P, De Weer A et al (2009) A novel and universal method for microRNA RT-qPCR data normalization. Genome Biol 10(6):R64. https://doi.org/10.1186/gb-2009-10-6-r64
D’Haene B, Mestdagh P, Hellemans J et al (2012) miRNA expression profiling: from reference genes to global mean normalization. Methods Mol Biol 822:261–272. https://doi.org/10.1007/978-1-61779-427-8_18
Schwarzenbach H, da Silva AM, Calin G et al (2015) Data normalization strategies for MicroRNA quantification. Clin Chem 61(11):1333–1342. https://doi.org/10.1373/clinchem.2015.239459
Broeders S, Huber I, Grohmann L et al (2014) Guidelines for validation of qualitative real-time PCR methods. Trends Food Sci Tech 37(2):115–126. https://doi.org/10.1016/j.tifs.2014.03.008
Kralik P, Ricchi M (2017) A basic guide to real time PCR in microbial diagnostics: definitions, parameters, and everything. Front Microbiol 8:108. https://doi.org/10.3389/fmicb.2017.00108
Vandesompele J, De Preter K, Pattyn F et al (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3(7):Research0034
Andersen CL, Jensen JL, Orntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64(15):5245–5250. https://doi.org/10.1158/0008-5472.CAN-04-0496
El-Khoury V, Pierson S, Kaoma T et al (2016) Assessing cellular and circulating miRNA recovery: the impact of the RNA isolation method and the quantity of input material. Sci Rep 6:19529. https://doi.org/10.1038/srep19529
McAlexander MA, Phillips MJ, Witwer KW (2013) Comparison of methods for miRNA extraction from plasma and quantitative recovery of RNA from cerebrospinal fluid. Front Genet 4:83. https://doi.org/10.3389/fgene.2013.00083
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Denk, J., Jahn, H. (2018). MicroRNA Profiling of Alzheimer’s Disease Cerebrospinal Fluid. In: Perneczky, R. (eds) Biomarkers for Alzheimer’s Disease Drug Development. Methods in Molecular Biology, vol 1750. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7704-8_6
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7704-8_6
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7703-1
Online ISBN: 978-1-4939-7704-8
eBook Packages: Springer Protocols