title: Analysis of microRNA expression and function.
abstract: Originally discovered in C. elegans, microRNAs (miRNAs) are small RNAs that regulate fundamental cellular processes in diverse organisms. MiRNAs are encoded within the genome and are initially transcribed as primary transcripts that can be several kilobases in length. Primary transcripts are successively cleaved by two RNase III enzymes, Drosha in the nucleus and Dicer in the cytoplasm, to produce ∼70 nucleotide (nt) long precursor miRNAs and 22 nt long mature miRNAs, respectively. Mature miRNAs regulate gene expression post-transcriptionally by imperfectly binding target mRNAs in association with the multiprotein RNA induced silencing complex (RISC). The conserved sequence, expression pattern, and function of some miRNAs across distinct species as well as the importance of specific miRNAs in many biological pathways have led to an explosion in the study of miRNA biogenesis, miRNA target identification, and miRNA target regulation. Many advances in our understanding of miRNA biology have come from studies in the powerful model organism C. elegans. This chapter reviews the current methods used in C. elegans to study miRNA biogenesis, small RNA populations, miRNA-protein complexes, and miRNA target regulation.
authors: Department of Biology, University of California San Diego, La Jolla, California, USA.
authors: Priscilla M
authors: PM
authors: Van Wynsberghe
authors: Department of Molecular, Cellular and Developmental Biology,Yale University, New Haven, Connecticut, USA.
authors: Shih-Peng
authors: SP
authors: Chan
authors: Department of Molecular, Cellular and Developmental Biology,Yale University, New Haven, Connecticut, USA.
authors: Frank J
authors: FJ
authors: Slack
authors: Department of Biology, University of California San Diego, La Jolla, California, USA.
authors: Amy E
authors: AE
authors: Pasquinelli
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D015870:Gene Expression; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D035683:MicroRNAs / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D018510:RNA, Helminth / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D017423:Sequence Analysis, RNA
chemical_list: D035683:MicroRNAs; D018510:RNA, Helminth; D016601:RNA-Binding Proteins