Hertel K.J. (Ed.) Spliceosomal Pre-mRNA Splicing. Methods and Protocols

Humana Press, 2014 — 427p. — (Methods in Molecular Biology 1126) — ISBN: 978-1-62703-980-2 (eBook), 978-1-62703-979-6 (Hardcover).Providing a guide to classical experimental approaches to decipher splicing mechanisms and experimental strategies that rely on novel multi-disciplinary approaches, Spliceosomal Pre-mRNA Splicing: Methods and Protocols describes the theory of alternative pre-mRNA splicing in seven introductory chapters and then introduces protocols and their theoretical background relevant for a variety of experimental research. These protocol chapters cover basic methods to detect splicing events, analyses of alternative pre-mRNA splicing in vitro and in vivo manipulation of splicing events and high-throughput and bioinformatic analyses of alternative splicing. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and practical, Spliceosomal Pre-mRNA Splicing: Methods and Protocols will aid newcomers and seasoned molecular biologists in understanding the fascinating world of alternative splicing with the ultimate goal of paving the way for many new discoveries to come.The Pre-mRNA Splicing Reaction
Diversity and Evolution of Spliceosomal Systems
Mechanisms of Spliceosomal Assembly
Alternative Pre-mRNA Splicing
Regulation of Alternative Pre-mRNA Splicing
Introduction to Cotranscriptional RNA Splicing
Chromatin and Splicing
Preparation of Splicing Competent Nuclear Extracts
Preparation of Yeast Whole Cell Splicing Extract
Efficient Splinted Ligation of Synthetic RNA Using RNA Ligase
In Vitro Assay of Pre-mRNA Splicing in Mammalian Nuclear Extract
Kinetic Analysis of In Vitro Pre-mRNA Splicing in HeLa Nuclear Extract
In Vitro Systems for Coupling RNAP II Transcription to Splicing and Polyadenylation
Isolation and Accumulation of Spliceosomal Assembly Intermediates
Complementation of U4 snRNA in S. cerevisiae Splicing Extracts for Biochemical Studies of snRNP Assembly and Function
Expression and Purification of Splicing Proteins from Mammalian Cells
Single Molecule Approaches for Studying Spliceosome Assembly and Catalysis
Cell-Based Splicing of Minigenes
Quantifying the Ratio of Spliceosome Components Assembled on Pre-mRNA
Antisense Methods to Modulate Pre-mRNA Splicing