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 centraleffectors 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-datemethods 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 thenecessary 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. 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. INDICE: 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, andTRBP Complexes.
- ISBN: 978-1-61779-045-4
- Editorial: Humana Press
- Encuadernacion: Cartoné
- Páginas: 315
- Fecha Publicación: 29/05/2011
- Nº Volúmenes: 1
- Idioma: Inglés