Summary
In this chapter, we provide a protocol for the production of transfected cell arrays in living mammalian cells on noncoated chambered coverglass for the systematic functional analyses of human genes by high-content screening microscopy. This method should facilitate drug target validation by small-interfering RNAs.
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References
Ziauddin, J. and Sabatini, D. M. (2001) Microarrays of cells expressing defined cDNAs. Nature 411, 107–110.
Erfle, H., Simpson, J. C., Bastiaens, P. I., and Pepperkok, R. (2004) siRNA cell arrays for high-content screening microscopy. BioTechniques 37, 454–462.
Liebel, U., Starkuviene, V., Erfle, H., et al. (2003) A microscope-based screening platform for large scale functional analysis in intact cells. FEBS Lett. 554, 394–398.
Starkuviene, V., Liebel, U., Simpson, J. C., et al. (2004) High-content screening microscopy identifies novel proteins with a putative role in secretory membrane traffic. Genome Res. 14, 1948–1956.
Simpson, J. C., Wellenreuther, R., Poustka, A., Pepperkok, R., and Wiemann, S. (2000) Systematic subcellular localization of novel proteins identified by large-scale cDNA sequencing. EMBO Rep. 1, 287–292.
Pepperkok, R., Girod, A., Simpson, J. C., and Rietdorf, J. (2000) Imunofluorescence microscopy, in Shepherd, P. and Dean, C., eds., Monoclonal Antibodies: A Practical Approach. Oxford University Press, New York, pp. 355–370.
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© 2007 Humana Press Inc.
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Erfle, H., Pepperkok, R. (2007). Production of siRNA and cDNA-Transfected Cell Arrays on Noncoated Chambered Coverglass for High-Content Screening Microscopy in Living Cells. In: Sioud, M. (eds) Target Discovery and Validation Reviews and Protocols. Methods in Molecular Biology™, vol 360. Humana Press. https://doi.org/10.1385/1-59745-165-7:155
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DOI: https://doi.org/10.1385/1-59745-165-7:155
Publisher Name: Humana Press
Print ISBN: 978-1-58829-656-6
Online ISBN: 978-1-59745-165-9
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