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Cystic Fibrosis: Methods and Protocols consolidates cutting-edge 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 produce multisystem pathology in both human and animal models.

Authors: Ed. Skach, William R

Table of Contents

1. CFTR Mutation Detection by Multiplex Heteroduplex (mHET) Analysis on MDE Gel (pp. 03-20)
   Zielenski, Julian; Aznarez, Isabel; Onay, Tuncer; Tzounzouris, John; Markiewicz, Danuta; Tsui, Lap-Chee
2. cDNA Microarrays for Pharmacogenomic Analysis of Cystic Fibrosis (pp. 21-30)
   Srivastava, Meera; Eidelman, Ofer; Pollard, Harvey B.
3. Natural Animal Models of Human Genetic Diseases (pp. 31-46)
   Wine, Jeffrey J.; Dean, Michael; Glavac, Damjan
4. Electrophysiological Approach to Studying CFTR (pp. 49-66)
   Fischer, Horst
5. Quantitative Analysis of ATP-Dependent Gating of CFTR (pp. 67-98)
   Powe, Allan; Zhou, Zhen; Hwang, Tzyh-Chang; Nagel, Georg
6. CFTR Regulation by Phosphorylation (pp. 99-110)
   Zhu, Tang; Hinkson, Deborah A. R.; Dahan, David; Evagelidis, Alexandra; Hanrahan, John W.
7. Transepithelial Measurements of Bicarbonate Secretion in Calu-3 Cells (pp. 111-128)
   Bridges, Robert J.
8. Transepithelial Impedance Analysis of Chloride Secretion (pp. 129-142)
   Singh, Ashvani K.; Singh, Sangeeta; Devor, Daniel C.; Frizzell, Raymond A.; Driessche, Willy van; Bridges, Robert J.
9. Studies of the Molecular Basis for Cystic Fibrosis Using Purified Reconstituted CFTR Protein (pp. 143-158)
   Kogan, Ilana; Ramjeesingh, Mohabir; Li, Canhui; Bear, Christine E.
10. Probing CFTR Channel Structure and Function Using the Substituted-Cysteine-Accessibility Method (pp. 159-174)
    Akabas, Myles H.
11. Methods for the Study of Intermolecular and Intramolecular Interactions Regulating CFTR Function (pp. 175-186)
    Naren, Anjaparavanda P.
12. Fluorescent Indicator Methods to Assay Functional CFTR Expression in Cells (pp. 187-196)
    Verkman, Alan S.; Jayaraman, Sujatha
13. Immunolocalization of CFTR in Intact Tissue and Cultured Cells (pp. 199-216)
    Marino, Christopher R.
14. Analysis of CFTR Trafficking and Polarization Using Green Fluorescent Protein and Confocal Microscopy (pp. 217-228)
    Moyer, Bryan D.; Stanton, Bruce A.
15. CFTR Folding and Maturation in Cells (pp. 229-244)
    Benharouga, Mohamed; Sharma, Manu; Lukacs, Gergely L.
16. Isolation of CFTR: Chaperone Complexes by Co-Immunoprecipitation (pp. 245-256)
    Meacham, Geoffrey C.; Cyr, Douglas M.
17. CFTR Expression and ER-Associated Degradation in Yeast (pp. 257-266)
    Zhang, Yimao; Michaelis, Susan; Brodsky, Jeffrey L.
18. Manipulating the Folding Pathway of deltaF508 CFTR Using Chemical Chaperones (pp. 267-276)
    Howard, Marybeth; Welch, William J.
19. CFTR Degradation and Aggregation (pp. 277-294)
    Corboy, Michael J.; Thomas, Philip J.; Wigley, W. Christian
20. In Vitro Reconstitution of CFTR Biogenesis and Degradation (pp. 295-310)
    Oberdorf, Jon; Skach, William R.
21. In Vitro CFTR Folding Assays (pp. 311-322)
    Stidham, Rhesa D.; Wigley, W. Christian; Thomas, Philip J.
22. Analysis of CFTR Endocytosis by Cell Surface Biotinylation (pp. 323-340)
    Weixel, Kelly; Bradbury, Neil A.
23. CFTR Regulation of ENaC (pp. 343-364)
    Donaldson, Scott H.; Poligone, Elaine G.; Stutts, M. Jackson
24. Yeast Two-Hybrid Identification and Analysis of Protein Interactions with CFTR (pp. 365-382)
    Raghuram, Viswanathan; Hallows, Kenneth R.; Foskett, J. Kevin
25. Biochemical Assays for Studying Indirect Interactions Between CFTR and the Cytoskeleton (pp. 383-394)
    Mohler, Peter J.; Kultgen, Patricia L.; Stutts, M. Jackson; Milgram, Sharon L.
26. CFTR-Associated ATP Transport and Release (pp. 395-406)
    Egan, Marie E.
27. Inflammatory Mediators in CF Patients (pp. 409-432)
    Hilliard, Jay B.; Konstan, Michael W.; Davis, Pamela B.
28. Bacterial Colonization and Infection in the CF Lung (pp. 433-446)
    Sagel, Scott D.; Dowell, Elaine B.; Accurso, Frank J.
29. Antimicrobial Peptides and Proteins in the CF Airway (pp. 447-464)
    Cole, Alexander M.; Ganz, Tomas
30. Bacterial-Epithelial Interactions (pp. 465-478)
    Bryan, Ruth; Prince, Alice
31. Thin-Film Measurements of Airway Surface Liquid Volume/Composition and Mucus Transport Rates In Vitro (pp. 479-492)
    Tarran, Robert; Boucher, Richard C.
32. Murine Models of CF Airway Infection and Inflammation (pp. 495-516)
    Chmiel, James F.; Konstan, Michael W.; Berger, Melvin
33. Analysis of Lipid Abnormalities in CF Mice (pp. 517-524)
    Freedman, Steven D.; Blanco, Paola G.; Shea, Julie C.; Alvarez, Juan G.
34. Bioelectric Measurement of CFTR Function in Mice (pp. 525-536)
    Grubb, Barbara R.
35. Xenograft Model of the CF Airway (pp. 537-550)
    Filali, Mohammed; Zhang, Yulong; Ritchie, Teresa C.; Engelhardt, John F.
36. Development of Conditionally Immortalized Epithelial Cell Lines from CF and Non-CF Mice (pp. 551-562)
    Cotton, Calvin U.
37. Technical Approaches to Analyze the In Vivo Ion Composition of Airway Surface Liquid (pp. 563-572)
    Zahm, Jean-Marie; Baconnais, Sonia; Balossier, Gerard; Puchelle, Edith
38. Design of Gene Therapy Clinical Trials in CF Patients (pp. 575-584)
    Curlee, Kimberly V.; Sorscher, Eric J.
39. Formulation of Synthetic Vectors for Cystic Fibrosis Gene Therapy (pp. 585-598)
    Marshall, John; Cheng, Seng H.
40. Adeno-Associated Viral Vectors for CF Gene Therapy (pp. 599-608)
    Flotte, Terence R.; Virella-Lowell, Isabel; Chesnut, Kye A.
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