Genetics of Cystic Fibrosis -- CFTR Mutation Detection by Multiplex Heteroduplex (mHET) Analysis on MDE Gel -- cDNA Microarrays for Pharmacogenomic Analysis of Cystic Fibrosis -- Natural Animal Models of Human Genetic Diseases -- CFTR structure and Function: -- Electrophysiological Approach to Studying CFTR -- Quantitative Analysis of ATP-Dependent Gating of CFTR -- CFTR Regulation by Phosphorylation -- Transepithelial Measurements of Bicarbonate Secretion in Calu-3 Cells -- Transepithelial Impedance Analysis of Chloride Secretion -- Studies of the Molecular Basis for Cystic Fibrosis Using Purified Reconstituted CFTR Protein -- Probing CFTR Channel Structure and Function Using the Substituted-Cysteine-Accessibility Method -- Methods for the Study of Intermolecular and Intramolecular Interactions Regulating CFTR Function -- Fluorescent Indicator Methods to Assay Functional CFTR Expression in Cells -- Immunolocalization of CFTR in Intact Tissue and Cultured Cells -- Analysis of CFTR Trafficking and Polarization Using Green Fluorescent Protein and Confocal Microscopy -- CFTR Folding and Maturation in Cells -- Isolation of CFTR -- CFTR Expression and ER-Associated Degradation in Yeast -- Manipulating the Folding Pathway of ?F508 CFTR Using Chemical Chaperones -- CFTR Degradation and Aggregation -- In Vitro Reconstitution of CFTR Biogenesis and Degradation -- In Vitro CFTR Folding Assays -- Analysis of CFTR Endocytosis by Cell Surface Biotinylation -- CFTR Regulation of ENaC -- Yeast Two-Hybrid Identification and Analysis of Protein Interactions with CFTR -- Biochemical Assays for Studying Indirect Interactions Between CFTR and the Cytoskeleton -- CFTR-Associated ATP Transport and Release -- Pathophysiology of Cystic Fibrosis -- Inflammatory Mediators in CF Patients -- Bacterial Colonization and Infection in the CF Lung -- Antimicrobial Peptides and Proteins in the CF Airway -- Bacterial-Epithelial Interactions -- Thin-Film Measurements of Airway Surface Liquid Volume/Composition and Mucus Transport Rates In Vitro -- Murine Models of CF Airway Infection and Inflammation -- Analysis of Lipid Abnormalities in CF Mice -- Bioelectric Measurement of CFTR Function in Mice -- Xenograft Model of the CF Airway -- Development of Conditionally Immortalized Epithelial Cell Lines from CF and Non-CF Mice -- Technical Approaches to Analyze the In Vivo Ion Composition of Airway Surface Liquid -- Novel Therapeutic Approaches for Cystic Fibrosis -- Design of Gene Therapy Clinical Trials in CF Patients -- Formulation of Synthetic Vectors for Cystic Fibrosis Gene Therapy -- Adeno-Associated Viral Vectors for CF Gene Therapy.

Since the cloning of the cystic fibrosis transmembrane conductance regulator (CFTR) nearly a decade ago, cystic fibrosis researchers, clinicians, and patients have come to rely increasingly on a diverse array of fundamental techniques to understand the molecular basis of this complex disease. Cystic Fibrosis Methods and Protocols consolidates a broad range of detailed and readily reproducible in vitro, cellular, and whole animal laboratory protocols into an indispensable resource. From electrophysiology and cell biology, to animal models and gene therapy, this comprehensive set of methods provides the step-by-step instructions needed for investigators to incorporate new approaches into their research programs. Specific protocols describe new techniques for diagnosis, in vitro methods for the expression and functional analysis of CFTR, novel biochemical and cellular systems to determine how mutations subvert CFTR function, and in vivo protocols to examine how CFTR dysfunction produces multisystem pathology in human and animal models. Comprehensive, multidisciplinary, and highly practical, Cystic Fibrosis Methods and Protocols makes accessible to today's cystic fibrosis investigator the powerful new scientific techniques required to investigate the basic science of the disease and to translate this into effective clinical solutions.