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Applications of Microscopy to Genetic Therapy of Cystic Fibrosis and Other Human Diseases

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Cell Imaging Techniques

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 319))

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Abstract

Gene therapy has become an extremely important and active field of biomedical research. Microscopy is an integral component of this effort. This chapter presents an overview of imaging techniques used in our facility in support of cystic fibrosis gene therapy research. Instrumentation used in these studies includes light and confocal microscopy, transmission electron microscopy, and scanning electron microscopy. Techniques outlined include negative staining, cryo-electron microscopy, three-dimentional reconstruction, enzyme cytochemistry, immunocytochemistry, and fluorescence imaging.

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References

  1. Rich, D. P., Anderson, M. P., Gregory, R. J., et al. (1990) Expression of cystic fibrosis transmembrane conductance regulator corrects defective chloride channel regulation in cystic fibrosis airway epithelial cells. Nature 347, 358–363.

    Article  PubMed  CAS  Google Scholar 

  2. Quinton. P. M. (1986) Missing Cl conductance in cystic fibrosis. Am. J. Physiol. 251, C649–C652.

    PubMed  CAS  Google Scholar 

  3. Smith, J. J., Travis, S. M., Greenberg, E. P., and Welsh, M. J. (1996) Cystic fibrosis airway epithelia fail to kill bacteria because of abnormal airway surface fluid. Cell 85, 229–236.

    Article  PubMed  CAS  Google Scholar 

  4. Romling, U., Fiedler, B., Bosshammer, J., et al. (1994) Epidemiology of chronic Pseudomonas aeruginosa infections in cystic fibrosis. J. Infect. Dis. 170, 1616–1621.

    PubMed  CAS  Google Scholar 

  5. Costerton, J. W., Stewart, P. S., and Greenberg, E. P. (1999) Bacterial biofilms: a common cause of persistent infections. Science 284, 1318–1322.

    Article  PubMed  CAS  Google Scholar 

  6. Riordan, J. R., Rommens, J. M., Kerem, B. S., et al. (1989) Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 245, 1066–1073.

    Article  PubMed  CAS  Google Scholar 

  7. Karp, P. H., Moninger, T. O., Weber, S. P., et al. (2002) An in vitro model of differentiated human airway epithelia. Methods for establishing primary cultures. Methods Mol. Biol. 188, 115–137.

    PubMed  Google Scholar 

  8. Fasbender, A. J., Zabner, J., and Welsh, M. J. (1995) Optimization of cationic lipid-mediated gene transfer to airway epithelia. Am. J. Physiol. 269, L45–L51.

    PubMed  CAS  Google Scholar 

  9. Fasbender, A. J., Marshall, J., Moninger, T. O., Grunst, T., Cheng, S., and Welsh, M. J. (1997) Effect of co-lipids in enhancing cationic lipid-mediated gene transfer in vitro and in vivo. Gene Ther. 4, 716–725.

    Article  PubMed  CAS  Google Scholar 

  10. Fasbender, A. J., Zabner, J., Chillon, M., et al. (1997) Complexes of adenovirus with polycationic polymers and cationic lipids increase the efficiency of gene transfer in vitro and in vivo. J. Biol. Chem. 272, 6479–6489.

    Article  PubMed  CAS  Google Scholar 

  11. Fasbender, A. J., Zabner, J., Zeiher, B. G., and Welsh, M. J. (1997) A low rate of cell proliferation and reduced DNA uptake limit cationic lipid-mediated gene transfer to primary cultures of ciliated airway epithelia. Gene Ther. 4, 1173–1180.

    Article  PubMed  CAS  Google Scholar 

  12. Zabner, J., Couture, L. A., Gregory, R. J., Graham, S. M., Smith, A. E., and Welsh, M. J. (1993) Adenovirus-mediated gene transfer transiently corrects the chloride transport defect in nasal epithelia of patients with cystic fibrosis. Cell 75, 207–216.

    Article  PubMed  CAS  Google Scholar 

  13. Wang, G., Zabner, J., Deering, C., et al (2000) Increasing epithelial junction permeability enhances gene transfer to airway epithelia In vivo. Am. J. Respir. Cell Mol. Biol. 22, 129–138.

    PubMed  Google Scholar 

  14. Ostedgaard, L. S., Zabner, J., Vermeer, D. W., et al. (2002) CFTR with a partially deleted R domain corrects the cystic fibrosis chloride transport defect in human airway epithelia in vitro and in mouse nasal mucosa in vivo. Proc. Natl. Acad. Sci. USA 99, 3093–3098.

    Article  PubMed  CAS  Google Scholar 

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© 2006 Humana Press Inc.

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Moninger, T.O., Nessler, R.A., Moore, K.C. (2006). Applications of Microscopy to Genetic Therapy of Cystic Fibrosis and Other Human Diseases. In: Taatjes, D.J., Mossman, B.T. (eds) Cell Imaging Techniques. Methods in Molecular Biology™, vol 319. Humana Press. https://doi.org/10.1007/978-1-59259-993-6_7

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  • DOI: https://doi.org/10.1007/978-1-59259-993-6_7

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-157-8

  • Online ISBN: 978-1-59259-993-6

  • eBook Packages: Springer Protocols

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