Main description:

This detailed book contains techniques to explore the unusual properties of the peculiar microsatellites known as trinucleotide repeats. Beginning with technical challenges raised by secondary structure-forming repeats and their propensity to contract and expand over time, the book continues with genetic screens in Saccharomyces cerevisiae, experimental systems to study trinucleotide repeat instability in human cells, dedicated experimental systems in appropriate cell types, as well as gene therapy approaches using the CRISPR-Cas family of endonucleases. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Trinucleotide Repeats: Methods and Protocols serves as a valuable aid to experts and newcomers alike who seek to investigate this fascinating and ever-expanding field of study.

Contents:

1. Experimenting with Trinucleotide Repeats: Facts and Technical Issues

Guy-Franck Richard

2. Fast Assays to Detect Interruptions in CTG.CAG Repeat Expansions

Stephanie Tome and Genevieve Gourdon

3. Tracking Expansions of Stable and Threshold Length Trinucleotide Repeat Tracts In Vivo and In Vitro Using Saccharomyces cerevisiae

Gregory M. Williams, Athena K. Petrides, Lata Balakrishnan, and Jennifer A. Surtees

4. Quantifying Replication Fork Progression at CTG Repeats by 2D Gel Electrophoresis

David Viterbo and Guy-Franck Richard

5. Genetic Assays to Study Repeat Fragility in Saccharomyces cerevisiae

Erica J. Polleys and Catherine H. Freudenreich

6. Genetic Screens to Study GAA/TTC and Inverted Repeat Instability in Saccharomyces cerevisiae

Wenying Guo and Kirill S. Lobachev

7. Monitoring Double-Strand Break Repair of Trinucleotide Repeats Using a Yeast Fluorescent Reporter Assay

Lucie Poggi, Bruno Dumas, and Guy-Franck Richard

8. Analysis of Trinucleotide Repeat Stability by Integration at a Chromosomal Ectopic Site

Rujuta Yashodhan Gadgil, S. Dean Rider, Todd Lewis, Joanna Barthelemy, and Michael Leffak

9. Experimental System to Study Instability of (CGG)n Repeats in Cultured Mammalian Cells

Artem V. Kononenko, Thomas Ebersole, and Sergei M. Mirkin

10. Assessing Triplet Repeat Expansions in Human SVG-A Cell Culture

Gregory M. Williams and Robert S. Lahue

11. The Isolation and Analysis of the CGG-Repeat Size in Male and Female Gametes from a Fragile X Mouse Model

Xiaonan Zhao, Huiyan Lu, Pradeep K. Dagur, and Karen Usdin

12. In Vitro Synthesis and RNA Structure Probing of CUG Triplet Repeat RNA

Remco T.P. van Cruchten and Derick G. Wansink

13. FISH Protocol for Myotonic Dystrophy Type 1 Cells

Arnaud F. Klein, Ludovic Arandel, Joelle Marie, and Denis Furling

14. Real Time Videomicroscopy and Semi-Automated Analysis of Brain Cell Culture Models of Trinucleotide Repeat Expansion Diseases

Sandra O. Braz, Diana M. Dinca, Genevieve Gourdon, and Mario Gomes-Pereira

15. Primary Cultures of Pure Embryonic Dorsal Root Ganglia Sensory Neurons as a New Cellular Model for Friedreich's Ataxia

Olivier Griso and Helene Puccio

16. GFP Reporters to Monitor Instability and Expression of Expanded CAG/CTG Repeats

Cinzia Cinesi, Bin Yang, and Vincent Dion

17. Gene Therapy for Huntington's Disease Using Targeted Endonucleases

Magdalena Dabrowska and Marta Olejniczak