Argonaute Proteins: Methods and Protocols

ISBN-13: 9781617790454 / Angielski / Twarda / 2011 / 344 str.

Tom C. Hobman; Thomas F. Duchaine

Opis

With the rapid proliferation of RNAi applications in basic and clinical sciences, the challenge has now become understanding how components of RNAi machinery function together in a regulated manner. Argonaute proteins are the central effectors of RNAi and are highly conserved among eukaryotes and some archaebacteria. These RNA-binding proteins use small guide RNAs to silence expression of genes at the mRNA and DNA levels. InArgonaute Proteins: Methods and Protocols, expert researchers in this burgeoning field provide detailed, up-to-date methods to study Argonaute protein functions and interactions in a wide variety of cell types ranging from yeast to mammalian systems, as well as in vitro. Written in the highly successfulMethods in Molecular Biology series format, chapters include brief 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.Practical and authoritative, Argonaute Proteins: Methods and Protocolsserves as a vital reference for both experienced and novice scientists approaching the vast complexities of RNAi research.

Recenzje

From the reviews:

"It can be fully appreciated the high quality of the book, the so well organized and sequentially logic presentation of the chapters. ... giving a very update and detailed presentation of the entire Argonaute family members, their evolutionary conservation in Eukaryotes, mechanisms of action, methods to study in vitro and in vivo ... . each biologist and physician, either involved in research or in clinical application will find irreplaceable this book. Hope the reader will get his own copy of this exciting book." (Manuela Monti, European Journal of Histochemistry, July, 2011)

Spis treści

1. Purification of Native Argonaute Complexes from the Fission Yeast Schizosaccharomyces pombe

            Shane M. Buker and Mohammad R. Motamedi

 

2. Chromatin Immuno-Precipitation in Fission Yeast

            Thomas A. Volpe and Jessica DeMaio

 

3. Biochemical Analyses of Endogenous Argonaute Complexes Immunopurified with Anti-Argonaute Monoclonal Antibodies

            Keita Miyoshi, Tomoko N. Okada, Haruhiko Siomi, and Mikiko C. Siomi

 

4. Mapping of Ago2-GW182 Functional Interactions

            Bing Yao, Songqing Li, Shangli Lian, Marvin J. Fritzler, and Edward K.L. Chan

 

5. Continuous Density Gradients to Study Argonaute and GW182 Complexes Associated with the Endocytic Pathway

            Derrick Gibbings

 

6. In vitro RISC Cleavage Assay

            Julia Stoehr and Gunter Meister

 

7. Native Gel Analysis for RISC Assembly

            Tomoko Kawamata and Yukihide Tomari

 

8. Purification and Assembly of Human Argonaute, Dicer, and TRBP Complexes

            Nabanita De and Ian J. MacRae

 

9. Detection of Human Dicer and Argonaute 2 Catalytic Activity

            Marjorie P. Perron, Patricia Landry, Isabelle Plante, and Patrick Provost

 

10. Imaging the Cellular Dynamics of Drosophila Argonaute Proteins

            Jing Li, Nima Najand, Wendy Long, and Andrew Simmonds

 

11. ­­­­Live Cell Imaging of Argonaute Proteins in Mammalian Cells

            Justin M. Pare, Joaquin Lopez-Orozco, and Tom C. Hobman

 

12. Reporter-Based Assays for Analyzing RNA Interference in Mammalian Cells

            Lydia V. McClure, Gil Ju Seo, and Christopher S. Sullivan

 

13. Artificial Tethering of Argonaute Proteins for Studying their Role in Translational Repression of Target mRNAs

            Stephanie Eckhardt, Emilia Szostak, Zhaolin Yang, and Ramesh Pillai

 

14. An Efficient System For Let-7 MicroRNA- And GW182 Protein-Mediated Deadenylation In vitro

            Marc R. Fabian, Yuri V. Svitkin, and Nahum Sonenberg

 

15. Cell-Free microRNA-Mediated Translation Repression in C. elegans

            Edlyn Wu and Thomas F. Duchaine

 

16. Argonaute Pull-Down and RISC Analysis Using 2'-O-Methylated Oligonucleotides Affinity Matrices

            Guillaume Jannot, Alejandro Vasquez-Rifo, and Martin J. Simard

 

17. Cloning Argonaute-Associated Small RNAs from C. elegans

            Weifeng Gu, Julie M. Claycomb, Pedro J Batista, Craig C. Mello, and Darryl Conte, Jr.

 

18. Immunoprecipitation of piRNPs and Directional, Next Generation Sequencing of piRNAs

            Yohei Kirino, Anastassios Vourekas, Eugene Khandros, and Zissimos Mourelatos

 

19. Generation of an Inducible Mouse ES Cell Line Deficient for Argonaute Proteins

            Hong Su and Xiaozhong Wang

 

20. Whole Cell Proteome Regulation by microRNAs Captured in a Pulsed SILAC Mass Spectrometry Approach

            Olivia Ebner and Matthias Selbach