Alternative pre-mRNA Splicing

Alternative pre-mRNA Splicing

Theory and Protocols
1. Aufl.

von: Stefan Stamm, Chris Smith, Reinhard Lührmann

99,99 €

Verlag: Wiley-Blackwell
Format: PDF
Veröffentl.: 09.01.2012
ISBN/EAN: 9783527636785
Sprache: englisch
Anzahl Seiten: 660

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This book was written for graduate and medical students, as well as clinicians and postdoctoral researchers. It describes the theory of alternative pre-mRNA splicing in<br> twelve introductory chapters and then introduces protocols and their theoretical background relevant for experimental research. These 43 practical chapters cover: Basic methods, Detection of splicing events, Analysis of alternative pre-mRNA splicing in vitro and in vivo, Manipulation of splicing events, and Bioinformatic analysis of alternative splicing.<br> A theoretical introduction and practical guide for molecular biologists, geneticists,clinicians and every researcher interested in alternative splicing.<br> <br> Website:
Companion Website:<br> <br> INTRODUCTION<br> Splicing in the RNA World (Emanuele Buratti, Maurizio Romano and Francisco E. Baralle)<br> RNPs, small RNAs, miRNAs (Michaela Beitzinger and Gunter Meister)<br> RNA Elements Involved in Splicing (William F. Mueller and Klemens J. Hertel)<br> A Structural Biology Perspective of Proteins Involved in Splicing Regulation (Antoine Clery and Frederic Allain)<br> The Spliceosome in Constitutive Splicing (Patrizia Fabrizio and Reinhard Luhrmann)<br> The Use of Saccharomyces cerevisiae to Study the Mechanism of pre-mRNA Splicing (Brian C. Rymond)<br> Challenges in Plant Alternative Splicing (Andrea Barta, Yamile Marquez and John WS Brown)<br> Alternative Splice Site Selection (Miguel B. Coelho and Christopher WJ Smith)<br> Integration of Splicing with Nuclear and Cellular Events (Aparna K. Sapra and Carla Neugebauer)<br> Splicing and Disease (Emanuele Buratti and Francisco E. Baralle)<br> From Bedside to Bench: How to Analyze a Splicing Mutation (Marco Baralle and Diana Baralle)<br> Analysis of Splicing Problems (Stefan Stamm)<br> BASIC METHODS <br> Ultracentrifugation in the Analysis and Purification by of Spliceosomes Assembled in Vitro<br> (Klaus Hartmuth, Maria A. van Santen and Reinhard Luhrmann)<br> Chemical Synthesis of RNA (Claudia Hobartner)<br> RNA Interference (siRNA, ShRNA) (Daphne S. Cabianca and Davide Gabellini)<br> Expression and Purification of Splicing Proteins (James Stevenin and Cyril F. Bourgeois)<br> Detection of RNA-Protein Complexes By Electrophoretic Mobility Shift Assay (Manli Shen and Michael G. Fried)<br> Functional Analysis of large Exonic Sequences through Iterative in Vivo Selection (Natalia N. Singh, Joonbae Seo and Ravindra N. Singh)<br> Identification of Splicing cis-Elements through an Ultra-Refined Antisense Microwalk (Ravindra N. Singh and Natalia N. Singh)<br> Genomic SELEX to Identify RNA Targets of Plant RNA Binding Proteins (Olga Bannikova and Andrea Barta)<br> DETECTION OF SPLICING EVENTS <br> Quantification of Alternative Splice Variants (Miriam LLorian and Christopher WJ Smith)<br> High-Throughput Analysis of Alternative Splicing by RT-PCR (Roscoe Klinck, Benoit Chabot and Sherif Abou Elela)<br> Monitoring Changes in Plant Alternative Splicing Events (Craig G. Simpson, Naeem Hasan Syed, Sujatha Manthri, John D. Fuller, Monika Maronova, Branislav Kusenda, Maria Kalyna, Andrea Barta and John WS Brown)<br> Array Analysis (Pierre de la Grange)<br> The CLIP Method to Study Protein-RNA Interactions in Intact Cells and Tissues (James Tollervey and Jernej Ule)<br> RNA: Protein Crosslinking and Immunoprecipitation in Yeast (CLIP) (Branislav Kusenda and Andrea Barta)<br> Identification of Proteins Bound to RNA (Emanuele Buratti)<br> Single Cell Detection of Splicing Events with Fluorescent Splicing Reporters (Hidehito Kuroyanagi, Akihide Takeuchi, Takayuki Nojima and Masatoshi Hagiwara)<br> ANALYSIS OF SPLICING IN VITRO <br> The Preparation of HeLa Cell Nuclear Extracts (Klaus Hartmuth, Maria A. van Santen, Tanja Rosel, Berthold Kastner and Reinhard Luhrmann)<br> In Vitro Splicing Assays (Akila Mayeda and Adrian R. Krainer)<br> Assembly of Spliceosomes in Vitro(Klaus Hartmuth, Maria A. van Santen, Peter Odenwalder and Reinhard Luhrmann)<br> Analysis of Site-Specific RNA-Protein Interactions (Nathalie Marmier-Gourrier, Audrey Vautrin, Christiane Branlant and Isabelle Behm-Ansmant)<br> Immunoprecipitation and Pull-Down of Nuclear Proteins<br> (Natalya Benderska, Chiranthani Sumanasekera and Stefan Stamm)<br> Analysis of Protein (RNA) Complexes by (Quantitative) Mass Spec Analysis (Carla Schmidt and Henning Urlaub)<br> ANALYSIS OF SPLICING IN VIVO <br> Fast cloning of splicing reporter minigenes (Zhaiyi Zhang, Amit Khanna and Stefan Stamm)<br> In Vivo Splicing Assays (Isabel Cristina Lopez Mejia and Jamal Tazi)<br> Coupled Promoter Splicing Systems (Manuel J. Mu?oz, Manuel de la Mata and Alberto R. Kornblihtt)<br> Stable Cell Lines With Splicing Reporters (Christian Kroun Damgaard, S?ren Lykke-Andersen and J?rgen Kjems)<br> Splicing Factor ChIP and ChRIP: Detection of Splicing and Splicing Factors at Genes by Chromatin Immunoprecipitation (Aparna K. Sapra, Fernando Carrillo Oesterreich, Marta Pabis, Imke Listerman, Nicole Bardehle and Karla M. Neugebauer)<br> Yeast Genetics to Investigate the Function of Core Pre-mRNA Splicing Factors (Raymond T. O?Keefe and Jean D. Beggs)<br> Analysis of HIV-1 RNA Splicing (Simon Duffy and Alan Cochrane)<br> In Vivo Analysis of Plant Intron Splicing (Craig G. Simpson, Michele Liney, Diane Davidson, Dominika Lewandowska, Maria Kalyna, Sean Chapman, Andrea Barta and John WS Brown)<br> Modificaton State-Specific Antibodies (Jordan B. Fishman, Olga Kelemen and Eric A. Berg)<br> Analysis of Alternative Splicing in Drosophila Genetic Mosaics (Shihuang Su, Diana O?Day, Shanzhi Wang and William Mattox)<br> MANIPULATION OF SPLICING EVENTS<br> Antisense Derivatives of U7 Small Nuclear RNA as Modulators of Pre-mRNA Splicing (Kathrin Meyer and Daniel Schumperli)<br> Screening for Alternative Splicing Modulators (Peter Stoilov)<br> Use of Oligonucleotides to Change Splicing (Annemieke Aartsma-Rus and Judith C.T. van Deutekom)<br> Changing Signals to the Spliceosome (Denise R. Cooper and Niketa A. Patel)<br> BIOINFORMATIC ANALYSIS OF SPLICING <br> Overview of Splicing Relevant Databases (Pierre de la Grange)<br> High Throughput Sequencing (Paolo Ribeca, Vincent Lacroix, Michael Sammeth and Roderic Guigo)<br> Identification of Splicing Factor Target Genes by High Throughput Sequencing<br> (Chaolin Zhang and Michael Q. Zhang)<br> Bioinformatic Analysis of Splicing Events (Zhaiyi Zhang and Stefan Stamm)<br> Analysis of Pre-mRNA Secondary Structures and Alternative Splicing (Michael Hiller)<br> Structure Prediction for Alternatively Spliced Proteins<br> (Lukasz Kozlowski, Jerzy Orlowski and Janusz M. Bujnicki)<br> Comparative Genomics Methods for the Prediction of Small RNA Binding Sites (Rym Kachouri-Lafond and Mihaela Zavolan)<br> <br>
"This book, prepared within the context of the EURASNET framework, is the first of its kind and, as such, an important piece of scientific literature, which will inspire more investigators to enter the rising field of alternative pre-mRNA splicing." (<i>Biotechnology Journal</i>, April 2012)
<b>Stefan Stamm</b> is Associate Professor in the Department for Molecular and Cellular Biochemistry at the University of Kentucky. He studied Biochemistry in Hannover (Germany) and did the practical work for his PhD as well as postdoctoral work at the Cold Spring Harbor Laboratory, NY. His research focuses on mechanisms and regulation of alternative splicing, with<br />the aim to apply the fi ndings to the Prader-Willi Syndrome and Spinal Muscular Atrophy.<br /><br /><b>Chris Smith</b> is Professor of RNA Molecular Biology in the Department of Biochemistry at the University of Cambridge. His PhD research in Biochemistry was carried out at the University of London, followed by postdoctoral research at Harvard Medical School. His research interests encompass the mechanisms, regulation and function of alternative splicing.<br /><br /><b>Reinhard Lührmann</b> is Director at the Max Planck Institute for Biophysical Chemistry in Göttingen where he is head of the Department of Cellular Biochemistry. He is an Honorary Professor at the University of Göttingen. He has studied chemistry at the University of Münster where he also received his PhD. He worked as a postdoctoral fellow at the Max Planck Institute<br />for Molecular Genetics in Berlin. His research focuses on the structure and mechanism of the spliceosome.
<p>All eukaryotic messenger RNAs are derived from precursors, the pre-mRNAs, by an extensive series of nuclear modifications. Most notably, the removal of introns and the joining of the remaining exons in the process of pre-mRNA splicing is essential to generate mRNAs that are suitable templates for translation.<br />The vast majority of human pre-mRNAs undergo alternative splicing, in which different RNA parts are either included or omitted in the mature mRNA. This process allows individual genes to generate multiple proteins and makes alternative pre-mRNA splicing a central element in gene regulation. Although pre-mRNA splicing occurs with high fidelity, an increasing number of diseases have been linked to defects that lead to mis-spliced mRNAs.</p> <p>This book was written for graduate and medical students, as well as clinicians and postdoctoral researchers. It describes the theory of alternative pre-mRNA splicing in twelve introductory chapters and then introduces protocols and their theoretical background relevant for experimental research. These 43 practical chapters cover: Basic methods, detection of splicing events, analysis of alternative pre-mRNA splicing in vitro and in vivo, manipulation of splicing events, and bioinformatic analysis of alternative splicing.</p> <p>A theoretical introduction and practical guide for molecular biologists, geneticists, clinicians and every researcher interested in alternative splicing.</p>

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