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High Throughput Screening of Gene Functions in Mammalian Cells Using Reversely Transfected Cell Arrays: Review And Protocol

[ Vol. 11 , Issue. 2 ]

Author(s):

Michael Sturzl, Andreas Konrad, Gaby Sander, Effi Wies, Frank Neipel, Elisabeth Naschberger, Simone Reipschlager, Nathalie Gonin-Laurent, Raymund E. Horch, Ulrich Kneser, Werner Hohenberger, Holger Erfle and Mathias Thurau   Pages 159 - 172 ( 14 )

Abstract:


Reversely transfected cell microarrays (RTCM) have been introduced as a method for parallel high throughput analysis of gene functions in mammalian cells. Hundreds to thousands of different recombinant DNA or RNA molecules can be transfected into different cell clusters at the same time on a single glass slide with this method. This allows either the simultaneous overexpression or - by using the recently developed RNA interference (RNAi) techniques - knockdown of a huge number of target genes. A growing number of sophisticated detection systems have been established to determine quantitatively the effects of the transfected molecules on the cell phenotype. Several different cell types have been successfully used for this procedure. This review summarizes the presently available knowledge on this technique and provides a laboratory protocol.

Keywords:

Reverse transfection, cell microarray, cell chip, apoptosis, proliferation, siRNA

Affiliation:

University of Erlangen-Nuremberg, Department of Surgery, Division of Molecular and Experimental Surgery, Schwabachanlage 10, D-91054 Erlangen, Germany.



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