Argonaute proteins : methods and protocols
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Bibliographic Information
Argonaute proteins : methods and protocols
（Methods in molecular biology / John M. Walker, series editor, v. 725）（Springer protocols）
Humana Press, c2011
- : hbk
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Includes bibliographical references and index
Description and Table of Contents
Description
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. In Argonaute 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 successful Methods in Molecular Biology (TM) 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 Protocols serves as a vital reference for both experienced and novice scientists approaching the vast complexities of RNAi research.
Table of Contents
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
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