Trinucleotide Repeat Protocols
(Sprache: Englisch)
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...
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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.
Klappentext zu „Trinucleotide Repeat Protocols “
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.
Inhaltsverzeichnis zu „Trinucleotide Repeat Protocols “
Mouse Models of Triplet Repeat Diseases.- Analysis of Triplet Pepeat DNAs and RNAs.- Analysis of Triplet Repeat Replication by Two-Dimensional Gel Electrophoresis.- Genetic Assays for Triplet Repeat Instability in Yeast.- Detection and Isolation of Trinucleotide Repeat Expansions Using the RED Method.- Analysis of Unstable Triplet Repeats Using Small-Pool Polymerase Chain Reaction.- Real-Time RT-PCR for CTG Repeat-Containing Genes.- Detection and Analysis of Polyglutamine-Containing Proteins and Their Aggregates.- Antibodies Against Huntingtin.- Using Antibodies to Analyze Polyglutamine Stretches.- Solubilization of Aggregates Formed by Expanded Polyglutamine Tract Expression in Cultured Cells.- Establishment of Animal and Cultured Cell Models for Trinucleotide Repeat Diseases.- Caenorhabditis elegans as a Model System for Triplet Repeat Diseases.- Monitoring Aggregate Formation in Organotypic Slice Cultures From Transgenic Mice.- The CGG Repeat and the FMR1 Gene.- Analysis of CTG Repeats Using DM1 Model Mice.- Lentiviral-Mediated Gene Transfer to Model Triplet Repeat Disorders.- Mouse Tissue Culture Models of Unstable Triplet Repeats.- In Vivo Analysis of Trinucleotide Repeat Diseases.- Neurotransmitter Receptor Analysis in Transgenic Mouse Models.- Chromatin Immunoprecipitation Technique for Study of Transcriptional Dysregulation in Intact Mouse Brain.- Techniques for Thick-Section Golgi Impregnation of Formalin-Fixed Brain Tissue.- Assessment of Impaired Proteasomal Function in a Cellular Model of Polyglutamine Diseases.- Assessment of In Vitro and In Vivo Mitochondrial Function in Friedreich's Ataxia and Huntington's Disease.- Triplet Repeats and DNA Repair.- Oxidative Damage in Huntington's Disease.
Autoren-Porträt
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,
Bibliographische Angaben
- 2010, XII, 342 Seiten, Maße: 15,2 x 22,9 cm, Kartoniert (TB), Englisch
- Herausgegeben: Yoshinori Kohwi
- Verlag: Springer, Berlin
- ISBN-10: 1617374466
- ISBN-13: 9781617374463
Sprache:
Englisch
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