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Intestinal Stem Cells pp 249–255Cite as

Specific Gene Expression in Lgr5+ Stem Cells by Using Cre-Lox Recombination

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2171))

Abstract

Intestinal stem cells are responsible for tissue renewal. The study of stem cell properties has become a major challenge in the field. We describe here a method based on Cre recombinase inducible lentivirus vectors that permits delivery of transgenes, either for overexpression or knockdown, in primary stem cells that can be cultured in an 3D intestinal organoid system. This method is an excellent approach for genetic manipulation and can complement in vivo transgenic experiments.

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References

  1. Barker N, van Es JH, Kuipers J, Kujala P, van den Born M, Cozijnsen M et al (2007) Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449(7165):1003–1007

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  2. Ordóñez-Morán P, Dafflon C, Imajo M, Nishida E, Huelsken J (2015) HOXA5 counteracts stem cell traits by inhibiting Wnt signaling in colorectal cancer. Cancer Cell 28(6):815–829

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  3. Chiacchiera F, Rossi A, Jammula S, Piunti A, Scelfo A, Ordóñez-Morán P et al (2016) Polycomb complex PRC1 preserves intestinal stem cell identity by sustaining Wnt/β-catenin transcriptional activity. Cell Stem Cell 18(1):91–103

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Acknowledgments

We would like to thank Didier Negre (École NS de Lyon, France) for the background vector SIV-GAE-SFFV. P.O-M was supported by EMBO and the University of Nottingham, UK.

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Authors and Affiliations

  1. Swiss Institute for Experimental Cancer Research, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland

    Pierre Dessen & Joerg Huelsken

  2. Division of Cancer & Stem Cells, School of Medicine, Biodiscovery Institute, Centre for Cancer Sciences, University of Nottingham, Nottingham, UK

    Paloma Ordóñez-Morán

Authors
  1. Pierre Dessen
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  2. Joerg Huelsken
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  3. Paloma Ordóñez-Morán
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Corresponding author

Correspondence to Paloma Ordóñez-Morán .

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Editors and Affiliations

  1. Division of Cancer and Stem Cells, School of Medicine, Biodiscovery Institute, Centre for Cancer Sciences, University of Nottingham, Nottingham, UK

    Paloma Ordóñez-Morán

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Dessen, P., Huelsken, J., Ordóñez-Morán, P. (2020). Specific Gene Expression in Lgr5+ Stem Cells by Using Cre-Lox Recombination. In: Ordóñez-Morán, P. (eds) Intestinal Stem Cells. Methods in Molecular Biology, vol 2171. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0747-3_16

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  • DOI: https://doi.org/10.1007/978-1-0716-0747-3_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0746-6

  • Online ISBN: 978-1-0716-0747-3

  • eBook Packages: Springer Protocols

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