Mirna Maturation: Methods and Protocols

ISBN-13: 9781493960163 / Angielski / Miękka / 2016 / 192 str.

Christoph Arenz

Opis

In miRNA Maturation: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used to study miRNA maturation. These included established methods such as fluorescent and non-fluorescent methods for homogenous assays of Dicer-mediated miRNA maturation or an in vivo assay for Drosha activity. Moreover, the volume also contains useful, but less-common methods that are hard to find elsewhere. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.Authoritative and practical, miRNA Maturation: Methods and Protocols seeks to widen the view on miRNA as biological mediator and potential drug target.

Spis treści

Part I

1. The Pathway of miRNA Maturation

Michael Sand

2. Micro RNA Maturation and Human Disease

Marlen Hesse and Christoph Arenz

3. Approaches to the Modulation of miRNA Maturation

Valerie T. Tripp, Jaclyn R. McKenna, and Douglas D. Young

 Part II

4. Expression Profiling of Components of the miRNA Maturation Machinery

Michael Sand and Marina Skrygan

5. Primary microRNA Processing Assay Reconstituted using Recombinant Drosha and DGCR8

Ian Barr and Feng Guo

6. In-vivo processing Assay Based on a Dual Luciferase Reporter System to Evaluate DROSHA Enzymatic Activity

Vera Bilan, Danilo Allegra, Florian Kuchenbauer, Daniel Mertens

7. Assaying Dicer-mediated miRNA Maturation by Means of Fluorescent Substrates

Marlen Hesse, Brian P. Davies and Christoph Arenz

8. A Fluorescence Correlation Spectroscopy-based Enzyme Assay for Human Dicer

Eileen Magbanua and Ulrich Hahn

9. Detection of Micro RNA Maturation Using Unmodified pre-miRNA and Branched Rolling Circle Amplification

Saskia Neubacher and Christoph Arenz

10. Identification of Inhibitors of microRNA Function from Small Molecule Screens

Colleen M. Connelly and Alexander Deiters

11. Quantitative RT-PCR Specific for Precursor and Mature miRNAs

Hannah Zöllner, Stephan A. Hahn and Abdelouahid Maghnouj.

12. Cellular MicroRNA Sensors Based on Luciferase Reporters

Colleen M. Connelly and Alexander Deiters

13. Inhibition of miRNA maturation by Peptide Nucleic Acids

Concetta Avitabile, Enrica Fabbri, Nicoletta Bianchi, Roberto Gambari and  Alessandra Romanelli

14. Molecular Methods for Validation of the Biological Activity of Peptide Nucleic Acids Targeting MicroRNAs

Eleonora Brognara, Enrica Fabbri, Nicoletta Bianchi, Alessia Finotti, Roberto Corradini,  and Roberto Gambari

15. Lentiviral Overexpression of miRNAs

Hannah Zöllner, Stephan A. Hahn and Abdelouahid Maghnouj