A collection of readily reproducible techniques for the study of mRNA processing and metabolism. These cutting-edge techniques range from cotranscriptional processing events that occur while the mNA is engaged with elongating RNA polymerase II, to in vivo and in vitro splicing and its biochemical analysis, and alternative splicing. Additional methods cover mRNA export, the recovery and analysis of mRNP complexes, cytoplasmic translation, mRNA degradation in vivo and in vitro, and the controversial concept of nuclear translation. Each proven protocol is described in step-by-step detail and contains a background introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding know pitfalls.mRNA processing is a key step in gene expression that effects all the proteins within the cell. In mRNA Processing and Metabolism: Methods and Protocols, world-renowned researchers bring together the latest techniques spanning the breadth of mRNA processing and metabolism. Drawing on recent advances in microscopy, whole genome sequencing, microarrays, mass spectrometry, fluorescent detection methodologies, and RNA interference, the authors offer readily reproducible methods for the cotranscriptional processing events that occur while the mRNA is engaged with elongating RNA polymerase II, with splicing and its biochemical analysis and with alternative splicing. Additional methods cover mRNA export, the recovery and analysis of mRNP complexes, cytoplasmic translation, mRNA degradation in vivo and in vitro, and the controversial concept of nuclear translation. A variety of model organisms are used, including yeast, Drosophila, Xenopus, mice, plants, and cultured mammalian cells. Each proven protocol is described in step-by-step detail and contains an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. §Timely and authoritative, mRNA Processing and Metabolism: Methods and Protocols provides a powerful collection of novel techniques that are not only effective, but also immediately applicable to current problems in many areas of biological research.
