title: Simple Methods to Investigate MicroRNA Induction in Response to Toll-Like Receptors.
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.
authors.affiliation: School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin 2, Ireland.|Hudson Institute of Medical Research, Monash University Faculty of Medicine, 27-31 Wright Street, Clayton, VIC 3168, Australia.
authors.forename: Victoria G
authors.initials: VG
authors.lastname: Lyons
authors.affiliation: School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin 2, Ireland. claire.mccoy@hudson.org.au.|Hudson Institute of Medical Research, Monash University Faculty of Medicine, 27-31 Wright Street, Clayton, VIC 3168, Australia. claire.mccoy@hudson.org.au.
authors.forename: Claire E
authors.initials: CE
authors.lastname: McCoy
mesh_terms: D000818:Animals; D015870:Gene Expression; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D008262:Macrophage Activation / Q000235:genetics / Q000276:immunology; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D051193:Toll-Like Receptors / Q000378:metabolism*; D059467:Transcriptome; D014162:Transfection
chemical_list: D035683:MicroRNAs; D051193:Toll-Like Receptors
keywords: BMDM; Bone marrow-derived macrophages; Luciferase assay; RT-PCR; TaqMan MicroRNA Array; Toll-like receptors; miRNA; miRNA inhibitor; miRNA mimic; microRNA