TY - BOOK AU - Hobman,Tom C. AU - Duchaine,Thomas F. ED - SpringerLink (Online service) TI - Argonaute Proteins: Methods and Protocols T2 - Methods in Molecular Biology, SN - 9781617790461 AV - QD431-431.7 U1 - 572.6 23 PY - 2011/// CY - Totowa, NJ PB - Humana Press, Imprint: Humana KW - Proteins  KW - Biotechnology KW - Protein Biochemistry KW - Chemical Bioengineering N1 - 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 N2 - 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 UR - https://doi.org/10.1007/978-1-61779-046-1 ER -