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A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture
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Biologie
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JoVE Journal Biologie
A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture

A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture

29,146 Views

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09:18 min

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August 11, 2014

DOI:

10.3791/51765-v

DOI:
10.3791/51765-v

•

09:18 min
August 11, 2014

•

5 Views
, , ,
1Department of Genetics,University of Cambridge, 2Wellcome Trust – Medical Research Council Stem Cell Institute,University of Cambridge

Summary

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This protocol explains primary Lgr5-positve organoid culture and the subsequent performance of retroviral transduction. This enables Cre-inducible overexpression or knockdown of the delivered transgene and allows functional studies to be carried out in the novel in vitro organotypic model system.

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Organoid CultureLgr5-positive Stem CellsEgfNogginR-Spondin3D Epithelial StructuresIn VitroIn VivoRetroviral TransductionFunctional GeneticsGene OverexpressionGene KnockdownSmall Intestinal EpitheliumPhenotypic AnalysisOrganotypic Model SystemGene Function

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Andersson-Rolf, A., Fink, J., Mustata, R. C., Koo, B. A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture. J. Vis. Exp. (90), e51765, doi:10.3791/51765 (2014).

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