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A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture
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A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture

A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture

DOI:
10.3791/51765-v

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09:18 min

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August 11, 2014

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

Chapters

  • 00:05Title
  • 01:17Crypt Isolation
  • 03:39Virus Production
  • 05:15Retroviral Transduction
  • 06:34Spinoculation and Plating
  • 08:01Results: Representative Organoid Images
  • 08:49Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

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

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