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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.
Cytology. --- Cell Biology. --- Cell biology --- Cellular biology --- Biology --- Cells --- Cytologists
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Two decades have passed since trinucleotide repeat expansion was first discovered in genes responsible for certain neurological diseases. Since then, new technologies have developed and innovative concepts have emerged, which may prove useful in devising therapeutic approaches to neurological diseases. Divided into six convenient sections, Trinucleotide Repeat Protocols, Second Edition covers a wide range of topics such as an overview of trinucleotide repeat diseases, synaptic plasticity, embryonic stem (ES) cell-related protocols with a focus on HD, RNA-related protocols, and analysis of epigenetic modification in fragile X syndrome. This edition focuses not only on direct analysis of trinucleotide repeat diseases but also on alternative approaches for the analysis of trinucleotide repeat diseases, with the hope that this will result in a better understanding of the mechanisms and future therapeutic prospects for treatment of these diseases. Written in the successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Trinucleotide Repeat Protocols, Second Edition seeks to serve researchers with its thorough methodologies on this expanding field.
Medicine. --- Gene expression. --- Molecular Medicine. --- Gene Expression. --- Genes --- Genetic regulation --- Health Workforce --- Expression
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