Main description:

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 (TM) 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.

Contents:

Part I. Introduction for Trinucleotide Repeat diseases

1. A Brief History of Triplet Repeat Disease

Helen Budworth and Cynthia McMurray

2. Huntington's Disease and Cell Therapies: Past, Present and Future

Chrystalina A. Antoniades and Colin Watts

Part II. Analysis of synaptic plasticity

3. Transcranial Two-Photon Imaging of Synaptic Structures in the Cortex of Awake Head-Restrained Mice

Guang Yang, Feng Pan, Paul C. Chang, Frank Gooden, and Wen-Biao Gan

4. In vivo Imaging of Synapse Plasticity in the Mouse Motor Cortex

Yi Zuo, Xinzhu Yu, Kelly Tennant, and Theresa Jones

Part III. Targeting on ES cells

5. Pluripotent Hybrid Stem Cells from Transgenic Huntington's Disease Monkey

Chuti Laowtammathron and Anthony W.S. Chan

6. Mesenchymal Stem Cells for Trinucleotide Repeat Disorders

Geralyn Annett, Gerhard Bauer, and Jan A. Nolta

Part IV. Targeting on RNA

7. Lentiviral-Mediated Gene Transfer of siRNAs for the Treatment of Huntington's Disease

Karine Cambon and Nicole Deglon

8. Purification of dFMR1-Containing Complexes Using Tandem Affinity Purification

Keita Miyoshi, Akiyo Ogino, Mikiko C. Siomi, and Haruhiko Siomi

9. Combined FISH and Immunofluorescent Staining Methods to Co-localize Proteins and mRNA in Neurons and Brain Tissue

Bin Ma and Naoko Tanese

Part V. Modification of Proteins and DNAs

10. Epigenetic Modifications of the FMR1 Gene

Elisabetta Tabolacci and Giovanni Neri

11. The CGG Repeat and the FMR1 Gene

Renate K. Hukema and Ben A. Oostra

12. Study of Oxidative Damage and Antioxidant Systems in Two Huntingtons Disease Rodent Models

Francisca Perez-Severiano, Sergio Montes-Lopez, Cristian Geronimo-Olvera, and Jose Segovia

13. Regulation of Ataxin-1 Phosphorylation and Its Impact on Biology

Sarita Lagalwar and Harry T. Orr

Part VI. Trinucleotide repat expansion and detection

14. Towards Understanding Region-Specificity of Triplet Repeat Diseases: Coupled Immunohistology and Mass Spectrometry Imaging

Virginia Platt, Do Yup Lee, Christie Canaria, Ken Frankel, Susan Bernstein, and Cynthia McMurray

15. Antibodies and Intrabodies Against Huntingtin: Production and Screening of Monoclonals and Single-Chain Recombinant Forms

Ali Khoshnan, Susan Ou, Jan Ko, and Paul H. Patterson

16. Cell Recovery from DM1 Transgenic Mouse Tissue to Study (CTG)n Instability and DM1 Pathogenesis

Judith R. Brouwer, Laurent Foiry, and Genevieve Gourdon

17. Markerless Modification of Trinucleotide Repeat Loci in BACs

Kellie A. Benzow and Michael D. Koob

18. DRPLA: Recent Advances in Research Using Transgenic Mouse Models

Kazushi Suzuki, Toshiya Sato, Mitsunori Yamada, Hitoshi Takahashi, and Shoji Tsuji