TY  - JOUR
A1  - Mousses, Spyro
A1  - Caplen, Natasha J.
A1  - Cornelison, Robert
A1  - Weaver, Don
A1  - Basik, Mark
A1  - Hautaniemi, Sampsa
A1  - Elkahloun, Abdel G.
A1  - Lotufo, Roberto A.
A1  - Choudary, Ashish
A1  - Dougherty, Edward R.
A1  - Suh, Ed
A1  - Kallioniemi, Olli
T1  - RNAi Microarray Analysis in Cultured Mammalian Cells
Y1  - 2003/10/01 
JF  - Genome Research 
JO  - Genome Research 
SP  - 2341 
EP  - 2347 
DO  - 10.1101/gr.1478703 
VL  - 13 
IS  - 10 
UR  - http://genome.cshlp.org/content/13/10/2341.abstract 
N2  - RNA interference (RNAi) mediated by small interfering RNAs (siRNAs) is a powerful new tool for analyzing gene knockdown phenotypes in living mammalian cells. To facilitate large-scale, high-throughput functional genomics studies using RNAi, we have developed a microarray-based technology for highly parallel analysis. Specifically, siRNAs in a transfection matrix were first arrayed on glass slides, overlaid with a monolayer of adherent cells, incubated to allow reverse transfection, and assessed for the effects of gene silencing by digital image analysis at a single cell level. Validation experiments with HeLa cells stably expressing GFP showed spatially confined, sequence-specific, time- and dose-dependent inhibition of green fluorescence for those cells growing directly on microspots containing siRNA targeting the GFP sequence. Microarray-based siRNA transfections analyzed with a custom-made quantitative image analysis system produced results that were identical to those from traditional well-based transfection, quantified by flow cytometry. Finally, to integrate experimental details, image analysis, data display, and data archiving, we developed a prototype information management system for high-throughput cell-based analyses. In summary, this RNAi microarray platform, together with ongoing efforts to develop large-scale human siRNA libraries, should facilitate genomic-scale cell-based analyses of gene function. 
ER  -