Cystic Fibrosis: Diagnosis and Protocols, Volume I: Approaches to Study and Correct CFTR Defects (Methods in Molecular Biology)

Despite the many milestones in cystic fibrosis (CF) research, progress towards curing the disease has been slow, and it is increasingly difficult to grasp and use the already wide and still growing range of diverse methods currently employed to study CF so as to understand it in its multidisciplinary nature.  Cystic Fibrosis: Diagnosis and Protocols aims to provide the CF research community and related researchers with a very wide range of high-quality experimental tools, as an easy way to grasp and use classical and novel methods applied to cystic fibrosis.  Volume I: Approaches to Study and Correct CFTR Defects focuses on the cystic fibrosis transmembrane conductance regulator (CFTR) and its expression, biogenesis, structure, and function in terms of the defects causing CF.  Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.  Comprehensive and practical, Cystic Fibrosis: Diagnosis and Protocols will provide readers with optimal working tools to address pressing questions in the best technical way, while helping all of us, as a research and clinical community, to move faster hand-in-hand toward unravelling the secrets of this challenging disorder and cure it. Part I: Strategies to Correct the Basic Defect in CF and Assess Efficacy in Human Clinical Trials   1. Introduction to Part I: The Relevance of CF Diagnostic Tools for Measuring Restoration of CFTR Function after Therapeutic Interventions in Human Clinical Trials             Kris De Boeck and Melissa Ashlock   2. High-Throughput Screening of Libraries of Compounds to Identify CFTR Modulators             Nicoletta Pedemonte, Olga Zegarra-Moran, and Luis J.V. Galietta   3. Repair of CFTR Folding Defects with Correctors that Function as Pharmacological Chaperones             Tip W. Loo and David M. Clarke   4. Use of Primary Cultures of Human Bronchial Epithelial Cells Isolated from Cystic Fibrosis Patients for the Pre-Clinical Testing of CFTR Modulators             Tim Neuberger, Bill Burton, Heather Clark, and Fredrick Van Goor   5. Design of Gene Therapy Trials in CF Patients             Jane C. Davies and Eric W.F.W. Alton   6. Nasal Potential Difference Measurements to Assess CFTR Ion Channel Activity             Steven M. Rowe, Jean-Paul Clancy, and Michael Wilschanski   7. Measurement of Ion Transport Function in Rectal Biopsies             Martin J. Hug, Nico Derichs, Inez Bronsveld, and Jean Paul Clancy   Part II: RNA Methods to Approach CFTR Expression   8. Introduction to Part II: RNA Methods to Approach CFTR Expression             Ann Harris   9. Quantification of CFTR Transcripts             Anabela S. Ramalho, Luka A. Clarke, and Margarida D. Amaral   10. Nonsense-Mediated mRNA Decay and Cystic Fibrosis             Liat Linde and Batsheva Kerem   11. Approaches to Study CFTR Pre-mRNA Splicing Defects             Elisa Goina, Eugenio Fernandez-Alanis, and Franco Pagani   12. Impact of MicroRNA in Normal and Pathological Respiratory Epithelia             Lisa Giovannini-Chami, Nathalie Grandvaux, Laure-Emmanuelle Zaragosi, Karine Robbe-Sermesant, Brice Marcet, Bruno Cardinaud, Christelle Coraux, Yves Berthiaume, Rainer Waldmann, Bernard Mari, and Pascal Barbry   13. Genomic Approaches to Studying CFTR Transcriptional Regulation             Christopher J. Ott and Ann Harris   Part III: CFTR Protein Biogenesis, Folding, Degradation, and Traffic   14. Introduction to Part III: Biochemical Methods to Study CFTR Protein             Margarida D. Amaral and Gergely L. Lukacs   15. Analysis of CFTR Folding and Degradation in Transiently Transfected Cells             Diane E. Grove, Meredith F. N. Rosser, Richard L. Watkins, and Douglas M. Cyr   16. In Vitro Methods for CFTR Biogenesis             Yoshihiro Matsumura, LeeAnn Rooney, and William R. Skach   17. Analysis of CFTR Interactome in the Macromolecular Complexes             Chunying Li and Anjaparavanda P. Naren   18. Methods to Monitor Cell Surface Expression and Endocytic Trafficking of CFTR in Polarized Epithelial Cells             Jennifer M. Bomberger, William B. Guggino, and Bruce A. Stanton   19. Segmental and Subcellular Distribution of CFTR in the Kidney             Franois Jouret, Pierre J. Courtoy, and Olivier Devuyst   20. Endocytic Sorting of CFTR Variants Monitored by Single Cell Fluorescence Ratio Image Analysis (FRIA) in Living Cells             Herve Barrire, Pirjo Apaja, Tsukasa Okiyoneda, and Gergely L. Lukacs   Part IV: CFTR Structure   21. Introduction to Part IV: Biophysical Methods to Approach CFTR Structure             Juan L. Mendoza, Andr Schmidt, and Philip J. Thomas   22. CFTR Three-Dimensional Structure             Robert C. Ford, James Birtley, Mark F. Rosenberg, and Liang Zhang   23. Molecular Modeling Tools and Approaches for CFTR and Cystic Fibrosis             Adrian W.R. Serohijos, Patrick H. Thibodeau, and Nikolay V. Dokholyan   24. Biochemical and Biophysical Approaches to Probe CFTR Structure             Andr Schmidt, Juan L. Mendoza, and Philip J. Thomas   25. NMR Spectroscopy to Study the Dynamics and Interactions of CFTR             Voula Kanelis, P. Andrew Chong, and Julie D. Forman-Kay   Part V: CFTR Function   26. Introduction to Part V: Assessment of CFTR Function             Karl Kunzelmann   27. Application of High-Resolution Single-Channel Recording to Functional Studies of Cystic Fibrosis Mutants             Zhiwei Cai, Yoshiro Sohma, Silvia G. Bompadre, David N. Sheppard, and Tzyh-Chang Hwang   28. Electrophysiological, Biochemical, and Bioinformatic Methods for Studying CFTR Channel Gating and Its Regulation             Lszl Csandy, Paola Vergani, Attila Gulys-Kovcs, and David C. Gadsby   29. CFTR Regulation by Phosphorylation             Rodrigo Alzamora, J. Darwin King, Jr., and Kenneth R. Hallows   30. How to Measure CFTR-Dependent Bicarbonate Transport: From Single Channels to the Intact Epithelium             Martin J. Hug, Lane L. Clarke, and Michael A. Gray