High throughput screening of gene functions in mammalian cells using reversely transfected cell arrays: review and protocol.
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.
Stürzl
Michael
M
Division of Molecular and Experimental Surgery, Department of Surgery, University of Erlangen-Nuremberg, Schwabachanlage 10, D-91054 Erlangen, Germany. michael.stuerzl@uk-erlangen.de
Konrad
Andreas
A
Sander
Gaby
G
Wies
Effi
E
Neipel
Frank
F
Naschberger
Elisabeth
E
Reipschläger
Simone
S
Gonin-Laurent
Nathalie
N
Horch
Raymund E
RE
Kneser
Ulrich
U
Hohenberger
Werner
W
Erfle
Holger
H
Thurau
Mathias
M
D000818:Animals; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology; D020868:Gene Silencing; D005796:Genes / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D048348:Reverse Transcription / Q000235:genetics*; D046888:Tissue Array Analysis / Q000379:methods*; D014162:Transfection / Q000379:methods*