Established leaders in trinucleotide repeat disease describe in step-by-step detail their best techniques for studying trinucleotide pathology at the molecular level. The protocols cover a variety of targets, ranging from DNA and RNA to proteins and whole animals, and focus not only on causal genes, but also on their consequent affected products, such as transcription factors, neurotransmitter receptors, proteasomes, and mitochondria/oxidation damage. Experimental systems employed include E. coli, yeast, C. elegans, mouse, and generally take a clinical point of view. The authors utilize a wide range of techniques, including gel electrophoresis, quantitative RT-PCR, immunological analysis, antibody usage and its applications, receptor assays using radioisotope handling, gene delivery by virus, brain cell and organotypic cultures, gender dependency, and neuron structure analysis.
Part I. Introduction Mouse Models of Triplet Repeat Diseases Gillian P. Bates and David G. Hay Part II. Analysis of Triplet Repeat DNAs and RNAs Analysis of Triplet Repeat Replication by Two-Dimensional Gel Electrophoresis Maria M. Krasilnikova and Sergei M. Mirkin Genetic Analysis for Triplet Repeat Instability in Yeast Michael J. Dixon, Saumitri Bhattacharyya, and Robert S. Lahue Detection and Isolation of Trinucleotide Repeat Expansions Using the RED Method Qiu-Ping Yuan and Martin Schalling Analysis of Unstable Triplet Repeats Using Small-Pool Polymerase Chain Reaction Mario Gomes-Pereira, Sanjay I. Bidichandani, and Darren G. Monckton Real-Time RT-PCR for CTG Repeat-Containing Genes Maria Eriksson Part III. Detection and Analysis of Polyglutamine-Containing Proteins and Their Aggregates Antibodies Against Huntingtin: Production and Screening of Monoclonals and Single-Chain Recombinant Forms Ali Khoshnan, Susan Ou, Jan Ko, and Paul H. Patterson Using Antibodies to Analyze Polyglutamine Stretches Elizabeth Brooks, Montserrat Arrasate, Kenneth Cheung, and Steven M. Finkbeiner Solubilization of Aggregates Formed by Expanded Polyglutamine Tract Expression in Cultured Cells Noriko Hazeki and Ichiro Kanazawa Part IV. Establishment of Animal and Cultured Cell Models for Trinucleotide Repeat Diseases Caenorhabditis elegans as a Model System for Triplet Repeat Diseases Cindy Voisine and Anne C. Hart Monitoring Aggregate Formation in Organotypic Slice Cultures From Transgenic Mice Donna L. Smith and Gillian P. Bates The CGG Repeat and the FMR1 Gene Violeta Stoyanova and Ben A. Oostra Analysis of CTG Repeats Using DM1 Model Mice Cedric Savouret, Claudine Junien, and Genevieve Gourdon Lentiviral-Mediated Gene Transfer to Model Triplet Repeat Disorders Etienne Regulier, Diana Zala, Patrick Aebischer, and Nicole Deglon Mouse Tissue Culture Models of Unstable Triplet Repeats Mario Gomes-Pereira and Darren G. Monckton Part V. In Vivo Analysis of Trinucleotide Repeat Diseases Neurotransmitter Receptor Analysis in Transgenic Mouse Models Caroline L. Benn, Laurie A. Farrell, and Jang-Ho J. Cha Chromatin Immunoprecipitation Technique for Study of Transcriptional Dysregulation in Intact Mouse Brain Melissa W. Braveman, Alice S. Chen-Plotkin, George J. Yohrling, and Jang-Ho J. Cha Techniques for Thick-Section Golgi Impregnation of Formalin-Fixed Brain Tissue Tracie L. Moss and William O. Whetsell, Jr. Assessment of Impaired Proteasomal Function in a Cellular Model of Polyglutamine Diseases Nihar Ranjan Jana and Nobuyuki Nukina Assessment of In Vitro and In Vivo Mitochondrial Function in Friedreich's Ataxia and Huntington's Disease Anthony Schapira and Raffaele Lodi Triplet Repeats and DNA Repair: Germ Cell and Somatic Cell Instability in Transgenic Mice Irina V. Kovtun, Craig Spiro, and Cynthia T. McMurray Oxidative Damage in Huntington's Disease Jose Segovia and Francisca Perez-Severiano Index
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