Full Description
This detailed volume collects chapters that seek to expand our knowledge of molecular events and extended molecular networks in gene regulation. The contents explore numerous aspects of miRNA biogenesis, a sophisticated series of events that assure a finely tuned regulation of miRNA expression and activity. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, miRNA Biogenesis: Methods and Protocols aims to contribute to the further development of knowledge and application of miRNA biogenesis, both in the basic research lab and in the clinic.
Contents
High-throughput characterization of primary microRNA transcripts.- Identifying pri-miRNA transcription start sites.- Metabolic pulse-chase RNA labelling for pri-miRNA processing dynamics.- In Vitro System for Coupling RNAP II Transcription to Primary microRNA Processing and a Three-way System for RNAP II Transcription/Splicing/microRNA Processing.- Purification of Microprocessor-associated factors.- Arresting pri-miRNA processing with Target Site Blockers.- MicroRNA analysis using the quantitative real-time PCR reaction.- MicroRNA analysis using next-generation sequencing.- Identification of microRNA precursor-associated proteins.- Analysis of 3' end modifications in (precursor-) microRNAs by high-throughput.- Uncovering the stability of mature miRNAs by 4-thio-uridine metabolic labeling.- Detection of microRNA-target interactions by Chimera PCR (ChimP).- High-quality overlapping paired-end reads for the detection of A-to-I editing on small RNA.- Targeting miRNA for therapeutics using a Micronome based method for identification of miRNA-mRNA pairs and validation of key regulator miRNA.- Method for detection of miRNAs in non-model organisms with unreported database.- Detection and Verification of Mammalian Mirtrons by Northern Blotting.- Detecting agotrons in Ago CLIPseq data.
