Abstract
In this chapter, we describe simple methods to investigate microRNA (miRNA) induction in response to lipopolysaccharide, the ligand for Toll-Like Receptor-4 activation. In brief, we demonstrate how to investigate global miRNA induction and/or repression in bone marrow-derived macrophages using TaqMan MicroRNA Arrays, followed by methods to measure individual miRNAs and target mRNA expression. Moreover, we explain step-by-step instructions on how to modulate endogenous miRNA expression through the use of miRNA inhibitors and mimics as well as highlight how miRNA modulation can be used to confirm mRNA targeting via Luciferase reporter assay. Moreover, these methods can be applied to whichever cell type and cellular function under investigation.
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Acknowledgements
The authors would like to acknowledge the Health Research Board (HRB) Ireland together with Marie Curie for funding the project from which the experiments included in this chapter are derived. The authors would also like to acknowledge their current funding from the National Health and Medical Research Council (NHMRC) Australia. The authors would like to thank Charlotte Nejad for her thorough editing of this manuscript.
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Lyons, V.G., McCoy, C.E. (2016). Simple Methods to Investigate MicroRNA Induction in Response to Toll-Like Receptors. In: McCoy, C. (eds) Toll-Like Receptors. Methods in Molecular Biology, vol 1390. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3335-8_11
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DOI: https://doi.org/10.1007/978-1-4939-3335-8_11
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