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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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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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.
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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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DOI: https://doi.org/10.1038/nmeth.1802
