Functional Genomics Assays to Study CFTR Traffic and ENaC Function

  • Joana Almaça
  • Shehrazade Dahimène
  • Nicole Appel
  • Christian Conrad
  • Karl Kunzelmann
  • Rainer Pepperkok
  • Margarida D. Amaral
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 742)

Abstract

As several genomes have been sequenced, post-genomic approaches like transcriptomics and proteomics, identifying gene products differentially expressed in association with a given pathology, have held promise both of understanding the pathways associated with the respective disease and as a fast track to therapy. Notwithstanding, these approaches cannot distinguish genes and proteins with mere secondary pathological association from those primarily involved in the basic defect(s). New global strategies and tools identifying gene products responsible for the basic cellular defect(s) in CF pathophysiology currently being performed are presented here. These include high-content screens to determine proteins affecting function and trafficking of CFTR and ENaC.

Key words

Cystic fibrosis CFTR secretory traffic ENaC high-content screens siRNA functional genomics 

Notes

Acknowledgements

This work is supported by TargetScreen2 (EU-FP6-LSH-2005-037365) grant. J.A. is recipient of Ph.D. fellowship SFRH/BD/29134/2006 (FCT, Portugal). The authors wish to thank Beate Neumann and Jutta Bulkescher (Advanced Light Microscopy Core Facility, EMBL) for their expert technical advice.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Joana Almaça
    • 1
    • 2
  • Shehrazade Dahimène
    • 1
  • Nicole Appel
    • 3
  • Christian Conrad
    • 3
  • Karl Kunzelmann
    • 4
  • Rainer Pepperkok
    • 3
  • Margarida D. Amaral
    • 1
    • 5
  1. 1.Faculty of Sciences, BioFiG-Centre for Biodiversity and Functional and Integrative GenomicsUniversity of LisboaLisboaPortugal
  2. 2.Department of Genetics, Centre of Human GeneticsNational Institute of HealthLisboaPortugal
  3. 3.EMBL-European Molecular Biology LaboratoryHeidelbergGermany
  4. 4.Department of PhysiologyUniversity of RegensburgRegensburgGermany
  5. 5.Department of Genetics, Centre of Human GeneticsNational Institute of HealthLisboaPortugal

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