Time-lapse microscopy-based genome wide RNAi screening in live human cells

Specific knock-down of gene expression by RNA interference is the method of choice to study gene function in human cells. A uniquely detailed phenotypic readout of such hypomorphs is possible by live cell microscopy of appropriate fluorescent reporter proteins. Here, I will present a fully automated...

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Bibliographic Details
Main Authors: Neumann, B., Held, M., Liebel, U., Erfle, H., Rogers, P., Pepperkok, R., Ellenberg, J.
Format: Conference Proceeding
Language:English
Subjects:
Bioinformatics
Cells (biology)
Fluorescence
Gene expression
Genomics
Humans
Interference
Microscopy
Proteins
RNA
Online Access:Request full text
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Summary:Specific knock-down of gene expression by RNA interference is the method of choice to study gene function in human cells. A uniquely detailed phenotypic readout of such hypomorphs is possible by live cell microscopy of appropriate fluorescent reporter proteins. Here, I will present a fully automated method for RNAi screens in cultured human cells, combining reverse transfection by siRNA cell arrays, automated time-lapse fluorescence microscopy and computational phenotype analysis by image processing. The strategy is illustrated using an automatically scored mitosis assay and provides an easily scalable platform that we currently use in genome-wide RNAi screens for several cellular functions
ISSN:1945-7928
1945-8452
DOI:10.1109/ISBI.2006.1624898