Argonaute Proteins
Humana Press Inc. (Verlag)
978-1-61779-045-4 (ISBN)
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™ 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.
Purification of Native Argonaute Complexes from the Fission Yeast Schizosaccharomyces pombe.- Chromatin Immuno-Precipitation in Fission Yeast.- Biochemical Analyses of Endogenous Argonaute Complexes Immunopurified with Anti-Argonaute Monoclonal Antibodies.- Mapping of Ago2-GW182 Functional Interactions.- Continuous Density Gradients to Study Argonaute and GW182 Complexes Associated with the Endocytic Pathway.- In vitro RISC Cleavage Assay.- Native Gel Analysis for RISC Assembly.- Purification and Assembly of Human Argonaute, Dicer, and TRBP Complexes.- Detection of Human Dicer and Argonaute 2 Catalytic Activity.- Imaging the Cellular Dynamics of Drosophila Argonaute Proteins.- Live Cell Imaging of Argonaute Proteins in Mammalian Cells.- Reporter-Based Assays for Analyzing RNA Interference in Mammalian Cells.- Artificial Tethering of Argonaute Proteins for Studying their Role in Translational Repression of Target mRNAs.- An Efficient System For Let-7 MicroRNA- And GW182 Protein-Mediated Deadenylation In vitro.- Cell-Free microRNA-Mediated Translation Repression in C. elegans.- Argonaute Pull-Down and RISC Analysis Using 2'-O-Methylated Oligonucleotides Affinity Matrices.- Cloning Argonaute-Associated Small RNAs from C. elegans.- Immunoprecipitation of piRNPs and Directional, Next Generation Sequencing of piRNAs.- Generation of an Inducible Mouse ES Cell Line Deficient for Argonaute Proteins.- Whole Cell Proteome Regulation by microRNAs Captured in a Pulsed SILAC Mass Spectrometry Approach.
| Reihe/Serie | Methods in Molecular Biology ; 725 |
|---|---|
| Zusatzinfo | XII, 344 p. |
| Verlagsort | Totowa, NJ |
| Sprache | englisch |
| Maße | 178 x 254 mm |
| Themenwelt | Naturwissenschaften ► Biologie ► Biochemie |
| Naturwissenschaften ► Biologie ► Mikrobiologie / Immunologie | |
| Technik ► Umwelttechnik / Biotechnologie | |
| ISBN-10 | 1-61779-045-1 / 1617790451 |
| ISBN-13 | 978-1-61779-045-4 / 9781617790454 |
| Zustand | Neuware |
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