MicroRNAs in Development Methods and Protocols

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™ 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.

1. In situ Detection of microRNAs in Animals

            Dylan Sweetman

2. Detection of microRNAs in Plants by In situ Hybridization

            Éva Várallyay and Zoltán Havelda

3. Detecting Animal and Plant microRNAs by Northern Blotting

            Sara López-Gomollón

4. Profiling microRNAs by Real-time PCR

            Nana Jacobsen, Ditte Andreasen, and Peter Mouritzen

5. Detection of Small RNAs and microRNAs using Deep Sequencing Technology

            Ericka R. Havecker

6. Detection of microRNAs in Prostate Cancer Cells by microRNA Array

            Xiaoqing Tang, Xiaohu Tang, Jozsef Gal, Natasha Kyprianou, Haining Zhu , and Guiliang Tang

7. microRNA Knock Down by Cholesterol Conjugated Antisense Oligos in Mouse Organ Culture  

            Sharon Kredo-Russo and Eran Hornstein

8. Protocols for Use of Homologous Recombination Gene-targeting to Produce microRNA Mutants in Drosophila

            Ya-Wen Chen, Ruifen Weng, and Stephen M. Cohen

9. Engineering Elements for Gene Silencing: The Artificial microRNAs Technology  

            Pablo Andrés Manavella and Ignacio Rubio-Somoza

10. Mimicry Technology: Suppressing Small RNA Activity in Plants

            Ignacio Rubio-Somoza and Pablo Andrés Manavella

11. Experimental Validation of MicroRNA Targets using a Luciferase Reporter System       

            Francisco E. Nicolas

12. Experimental Identification of microRNA Targets by Immunoprecipitation of Argonaute Protein Complexes

13. Comprehensive Identification of miRNA Target Sites in Live Animals       

            Dimitrios G. Zisoulis , Gene W. Yeo, and Amy E. Pasquinelli

14. Target Validation of Plant microRNAs

            César Llave, José Manuel Franco-Zorrilla, Roberto Solano, and Daniel Barajas

15. A High-throughput Sequencing-based Methodology to Identify All Uncapped and Cleaved RNA Molecules in Eukaryotic Genomes

            Matthew W. Endres, Rebecca T. Cook, and Brian D. Gregory