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Controlled gene expression in primary Lgr5 organoid cultures

Nature Methods volume 9, pages 81–83 (2012)Cite this article

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Abstract

The study of gene function in endodermal epithelia such as of stomach, small intestine and colon relies heavily on transgenic approaches. Establishing such animal models is laborious, expensive and time-consuming. We present here a method based on Cre recombinase–inducible retrovirus vectors that allows the conditional manipulation of gene expression in primary mouse organoid culture systems.

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Figure 1: An inducible organoid transduction system.
Figure 2: Notch pathway inhibition in intestinal organoids.

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References

  1. Mouse Genome Sequencing Consortium. et al. Nature 420, 520–562 (2002).

  2. Kurth, R. & Bannert, N. Retroviruses Molecular Biology, Genomics and Pathogenesis (Caister Academic Press, 2010).

  3. Sato, T. et al. Nature 459, 262–265 (2009).

    Article  CAS  Google Scholar 

  4. Sato, T. et al. Gastroenterology 141, 1762–1772 (2011).

    Article  CAS  Google Scholar 

  5. Barker, N. et al. Cell Stem Cell 6, 25–36 (2010).

    Article  CAS  Google Scholar 

  6. Glaser, S., Anastassiadis, K. & Stewart, A.F. Nat. Genet. 37, 1187–1193 (2005).

    Article  CAS  Google Scholar 

  7. Barker, N. et al. Nature 449, 1003–1007 (2007).

    Article  CAS  Google Scholar 

  8. Wu, Y., Melton, D.W., Zhang, Y. & Hornsby, P.J. J. Biomed. Biotechnol. 2009, 901079 (2009).

    PubMed  PubMed Central  Google Scholar 

  9. Sato, T. et al. Nature 469, 415–418 (2011).

    Article  CAS  Google Scholar 

  10. de Lau, W. et al. Nature 476, 293–297 (2011).

    Article  CAS  Google Scholar 

  11. van Es, J.H. et al. Nature 435, 959–963 (2005).

    Article  CAS  Google Scholar 

  12. Chung, K.H. et al. Nucleic Acids Res. 34, e53 (2006).

    Article  Google Scholar 

  13. Stern, P. et al. Proc. Natl. Acad. Sci. USA 105, 13895–13900 (2008).

    Article  CAS  Google Scholar 

  14. VanDussen, K.L. & Samuelson, L.C. Dev. Biol. 346, 215–223 (2010).

    Article  CAS  Google Scholar 

  15. Weng, A.P. et al. Mol. Cell. Biol. 23, 655–664 (2003).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank L. Kaaij, P. Tetteh, H. Begthel and S. van den Brink for experimental help. Rosa-CreERT2 mice were donated by A. Smith (Wellcome Trust Centre for Stem Cell Research, University of Cambridge) and DN-MAML1 by J. Aster (Harvard Medical School). This work was funded in part by grants from the European Research Council, EU/232814-StemCeLLMark and the National Research Foundation of Korea, NRF-2011-357-C00093 (B.-K.K.), The Centre van Biomedical Genetic (D.E.S.), The Dutch Cancer Society, KWF/Hubr2007-3956 (T.S.), the EU FP7 Tornado-KBBE-222720 (W.K.), EMBO long-term fellowship (LTF to H.F.F.), the EU Marie Curie Fellowships, EU/236954-ICSC LGR5 (M.H.), Ti Pharma/T3-106 (J.H.E.) and the KNAW/3V-fund.

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Author notes

  1. Bon-Kyoung Koo and Daniel E Stange: These authors contributed equally to this work.

Authors and Affiliations

  1. Hubrecht Institute for Developmental Biology and Stem Cell Research, and University Medical Centre, Utrecht, The Netherlands

    Bon-Kyoung Koo, Daniel E Stange, Toshiro Sato, Wouter Karthaus, Henner F Farin, Meritxell Huch, Johan H van Es & Hans Clevers

  2. Department of Gastroenterology, School of Medicine, Keio University, Tokyo, Japan

    Toshiro Sato

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  1. Bon-Kyoung Koo
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  2. Daniel E Stange
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  3. Toshiro Sato
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  4. Wouter Karthaus
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  5. Henner F Farin
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  6. Meritxell Huch
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  7. Johan H van Es
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  8. Hans Clevers
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Contributions

B.-K.K., D.E.S. and H.C. conceived the project and wrote the manuscript. B.-K.K., T.S. and M.H. developed the infection protocol and optimized the culture conditions. B.-K.K.,D.E.S. and H.F.F. designed and constructed retroviral vectors. B.-K.K. and J.H.v.E. established the organoids from CreERT2-transgenic mice. W.K. performed the lentiviral transduction.

Corresponding author

Correspondence to Hans Clevers.

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The authors declare no competing financial interests.

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Supplementary Figures 1–5 and Supplementary Table 1 (PDF 3709 kb)

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Koo, BK., Stange, D., Sato, T. et al. Controlled gene expression in primary Lgr5 organoid cultures. Nat Methods 9, 81–83 (2012). https://doi.org/10.1038/nmeth.1802

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