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Trinucleotide Repeat Protocols

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Trinucleotide repeats are relatively common in the human genome. These simple repeats have received much attention since epoch-mak... Weiterlesen
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Beschreibung

Trinucleotide repeats are relatively common in the human genome. These simple repeats have received much attention since epoch-making discoveries were made that particular trinucleotide repeats are expanded in the causal genes of human hereditary neurological disorders. For example, the CGG repeat is expanded in fragile X syndrome at the 5' untranslated region (UTR) of its causal gene. In myotonic dystrophy, it is the CTG repeat that is expanded at the 3' UTR of its causal gene. The CAG repeat was also found expanded in coding regions of the genes responsible for X-linked spinal and bulbar muscular atrophy, Huntington's disease, spinocerebellar ataxia, and other disorders. On the other hand, expansion of the GAA repeat was identified in the intron of the gene responsible for the Friedreich's ataxia. For these trinucleotide repeat diseases, the longer the trinucleotide expansion, the earlier the age of onset and the more severe the syndrome. Thus, these findings that showed the intriguing link between a particular trinucleotide expansion and its associated neurological disorders have led to a new field of intensive study. Active research addressing the underlying mechanisms for trinucleotide repeat diseases has employed various approaches ranging from DNA biochemistry to animal models for the diseases. In particular, animal models for the triplet repeat diseases have provided excellent resources not only for understanding the mechanisms but also for exploring therapeutic interventions.

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,

Klappentext
The discovery that trinucleotide repeats significantly influence the age of onset and severity of a variety of hereditary neurological disorders has opened the door to a deeper understanding of the disease mechanisms involved, as well as to a more productive search for novel therapeutic interventions. In Trinucleotide Repeat Protocols, 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 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. Each protocol follows the successful Methods in Molecular Biology™ series format, offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and highly practical, Trinucleotide Repeat Protocols offers neuroscientists powerful tools to elucidate both normal brain function and the mechanisms of hereditary neurological disease, as well as to develop the next generation of therapies for neuronal genetic diseases.

Inhalt
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 Mário 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 Cédric Savouret, Claudine Junien, and Geneviève Gourdon Lentiviral-Mediated Gene Transfer toModel Triplet Repeat Disorders Etienne Régulier, Diana Zala, Patrick Aebischer, and Nicole Déglon Mouse Tissue Culture Models of Unstable Triplet Repeats Mário 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 José Segovia and Francisca Pérez-Severiano Index

Produktinformationen

Titel: Trinucleotide Repeat Protocols
Editor:
EAN: 9781588292438
ISBN: 978-1-58829-243-8
Format: Fester Einband
Herausgeber: Springer, Berlin
Genre: Biologie
Anzahl Seiten: 342
Gewicht: 1g
Größe: H229mm x B229mm x T152mm
Jahr: 2004
Untertitel: Englisch
Auflage: 2004

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