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Trinucleotide Repeat Protocols : Methods in Molecular Biology - Yoshinori Kohwi

Trinucleotide Repeat Protocols

Methods in Molecular Biology

By: Yoshinori Kohwi (Editor)

Hardcover Published: 18th June 2004
ISBN: 9781588292438
Number Of Pages: 342

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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.

Prefacep. v
Contributorsp. ix
Introduction
Mouse Models of Triplet Repeat Diseasesp. 3
Analysis of Triplet Repeat DNAs and RNAs
Analysis of Triplet Repeat Replication by Two-Dimensional Gel Electrophoresisp. 19
Genetic Analysis for Triplet Repeat Instability in Yeastp. 29
Detection and Isolation of Trinucleotide Repeat Expansions Using the RED Methodp. 47
Analysis of Unstable Triplet Repeats Using Small-Pool Polymerase Chain Reactionp. 61
Real-Time RT-PCR for CTG Repeat-Containing Genesp. 77
Detection and Analysis of Polyglutamine-Containing Proteins and Their Aggregates
Antibodies Against Huntingtin: Production and Screening of Monoclonals and Single-Chain Recombinant Formsp. 87
Using Antibodies to Analyze Polyglutamine Stretchesp. 103
Solubilization of Aggregates Formed by Expanded Polyglutamine Tract Expression in Cultured Cellsp. 129
Establishment of Animal and Cultured Cell Models for Trinucleotide Repeat Diseases
Caenorhabditis elegans as a Model System for Triplet Repeat Diseasesp. 141
Monitoring Aggregate Formation in Organotypic Slice Cultures From Transgenic Micep. 161
The CGG Repeat and the FMR1 Genep. 173
Analysis of CTG Repeats Using DM1 Model Micep. 185
Lentiviral-Mediated Gene Transfer to Model Triplet Repeat Disordersp. 199
Mouse Tissue Culture Models of Unstable Triplet Repeatsp. 215
In Vivo Analysis of Trinucleotide Repeat Diseases
Neurotransmitter Receptor Analysis in Transgenic Mouse Modelsp. 231
Chromatin Immunoprecipitation Technique for Study of Transcriptional Dysregulation in Intact Mouse Brainp. 261
Techniques for Thick-Section Golgi Impregnation of Formalin-Fixed Brain Tissuep. 277
Assessment of Impaired Proteasomal Function in a Cellular Model of Polyglutamine Diseasesp. 287
Assessment of In Vitro and In Vivo Mitochondrial Function in Friedreich's Ataxia and Huntington's Diseasep. 293
Triplet Repeats and DNA Repair: Germ Cell and Somatic Cell Instability in Transgenic Micep. 309
Oxidative Damage in Huntington's Diseasep. 321
Indexp. 335
Table of Contents provided by Rittenhouse. All Rights Reserved.

ISBN: 9781588292438
ISBN-10: 1588292436
Series: Methods in Molecular Biology
Audience: Professional
Format: Hardcover
Language: English
Number Of Pages: 342
Published: 18th June 2004
Publisher: Humana Press Inc.
Country of Publication: US
Dimensions (cm): 23.5 x 15.5  x 2.54
Weight (kg): 0.69