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Hobman, Tom C. (Herausgeber); Duchaine, Thomas F. (Herausgeber)

Argonaute Proteins Methods and Protocols (Gebundene Ausgabe)

Reihe: Methods in Molecular Biology 725

Springer-Verlag GmbH, Humana Press, Mai 2011

344 S. - Sprache: Englisch - 63 schwarz-weiße und 2 farbige Abbildungen, 3 schwarz-weiße Tabellen - 264x184x25 mm Book

ISBN: 1617790451 EAN: 9781617790454

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.

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-Aids researchers in examining these versatile proteins in a wide variety of cell types-Features practical, step-by-step protocols which open the door to future progress in the field of molecular therapeutics-Contains expert tips and vital implementation advice to ensure success


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


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)

ISBN 1-617790-45-1, ISBN 1-61-779045-1, ISBN 1-61779-045-1, ISBN 1-617-79045-1, ISBN 1-6177-9045-1

ISBN 978-1-617790-45-4, ISBN 978-1-61-779045-4, ISBN 978-1-61779-045-4, ISBN 978-1-617-79045-4, ISBN 978-1-6177-9045-4

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