MicroRNAs in Development

Since the discovery of microRNAs, developmental biologists have striven to understand the role of miRNAs in development and disease. MicroRNAs in Development: Methods and Protocols collects contributions from expert researchers in order to provide practical guidelines to this complex study. Divided into three convenient sections, this detailed volume covers various techniques to detect and profile miRNA expression, followed by protocols to manipulate the activity of miRNAs in various organisms, and it concludes with a section that outlines different methods to identify and validate miRNA targets in animals and plants. Written in the highly successful Methods in Molecular Biology(TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, MicroRNAs in Development: Methods and Protocols serves as a practical guide for scientists of all backgrounds and conveys the appropriate sense of fascination associated with this vital field of research.

In situ Detection of microRNAs in Animals.- Detection of microRNAs in Plants by In situ Hybridization.- Detecting Animal and Plant microRNAs by Northern Blotting.- Profiling microRNAs by Real-time PCR.- Detection of Small RNAs and microRNAs using Deep Sequencing Technology.- Detection of microRNAs in Prostate Cancer Cells by microRNA Array.- microRNA Knock Down by Cholesterol Conjugated Antisense Oligos in Mouse Organ Culture.- Protocols for Use of Homologous Recombination Gene-targeting to Produce microRNA Mutants in Drosophila.- Engineering Elements for Gene Silencing: The Artificial microRNAs Technology.- Mimicry Technology: Suppressing Small RNA Activity in Plants.- Experimental Validation of MicroRNA Targets using a Luciferase Reporter System.- Experimental Identification of microRNA Targets by Immunoprecipitation of Argonaute Protein Complexes.- Comprehensive Identification of miRNA Target Sites in Live Animals.- Target Validation of Plant microRNAs.- A High-throughput Sequencing-based Methodology to Identify All Uncapped and Cleaved RNA Molecules in Eukaryotic Genomes