Abstract
Using RNAi interference (RNAi), it is possible to study the effect of specific gene knockdowns in mammalian cells. In this protocol we present the automated preparation of “ready to transfect” multiwell plates and cell arrays, on which cells can be grown which are then reversely transfected with one type of siRNA in every individual well or spot. Additionally, different microscope types for screening approaches are compared and considerations about the information workflow are made.
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Acknowledgment
We like to thank Michael Berthold, FB Informatik und Informationswissenschaften, Uni Konstanz, and the whole KNIP-Team for image analysis support. The ViroQuant-CellNetworks RNAi screening facility is supported by the Federal Ministry of Education and Research (BMBF) and funded by the FORSYS program ViroQuant (Project 0313923) and by CellNetworks-Cluster of Excellence (EXC81).
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Gunkel, M., Beil, N., Beneke, J., Reymann, J., Erfle, H. (2015). Fluorescence Microscopy-Based RNA Interference Screening. In: Verveer, P. (eds) Advanced Fluorescence Microscopy. Methods in Molecular Biology, vol 1251. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2080-8_4
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DOI: https://doi.org/10.1007/978-1-4939-2080-8_4
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2079-2
Online ISBN: 978-1-4939-2080-8
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