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Tamoxifen-independent recombination of reporter genes limits lineage tracing and mosaic analysis using CreER(T2) lines
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Vascular Biology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Vascular Biology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Vascular Biology.
Uppsala Univ, Dept Immunol Genet & Pathol, Rudbeck Lab, Dag Hammarskjoldsvag 20, S-75185 Uppsala, Sweden;Karolinska Inst, ICMC, Dept Med Huddinge, Novum, Blickagangen 6, S-14157 Huddinge, Sweden.
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2019 (English)In: Transgenic research, ISSN 0962-8819, E-ISSN 1573-9368Article in journal (Refereed) Epub ahead of print
Abstract [en]

The CreER(T2)/loxP system is widely used to induce conditional gene deletion in mice. One of the main advantages of the system is that Cre-mediated recombination can be controlled in time through Tamoxifen administration. This has allowed researchers to study the function of embryonic lethal genes at later developmental timepoints. In addition, CreER(T2) mouse lines are commonly used in combination with reporter genes for lineage tracing and mosaic analysis. In order for these experiments to be reliable, it is crucial that the cell labeling approach only marks the desired cell population and their progeny, as unfaithful expression of reporter genes in other cell types or even unintended labeling of the correct cell population at an undesired time point could lead to wrong conclusions. Here we report that all CreER(T2) mouse lines that we have studied exhibit a certain degree of Tamoxifen-independent, basal, Cre activity. Using Ai14 and Ai3, two commonly used fluorescent reporter genes, we show that those basal Cre activity levels are sufficient to label a significant amount of cells in a variety of tissues during embryogenesis, postnatal development and adulthood. This unintended labelling of cells imposes a serious problem for lineage tracing and mosaic analysis experiments. Importantly, however, we find that reporter constructs differ greatly in their susceptibility to basal CreER(T2) activity. While Ai14 and Ai3 easily recombine under basal CreER(T2) activity levels, mTmG and R26R-EYFP rarely become activated under these conditions and are therefore better suited for cell tracking experiments.

Place, publisher, year, edition, pages
2019.
Keywords [en]
CreER(T2), Cre, loxP system, Lineage tracing, Mosaic analysis, Tamoxifen-independent recombination, Reporter-gene
National Category
Cell and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-397671DOI: 10.1007/s11248-019-00177-8ISI: 000494238200001PubMedID: 31641921OAI: oai:DiVA.org:uu-397671DiVA, id: diva2:1373984
Funder
Swedish Cancer Society, CAN2015/771Swedish Cancer Society, CAN 2016/535Swedish Research Council, VR2015-00550Swedish Research Council, 542-2014-3535EU, European Research Council, 2011-294556EU, European Research Council, ERC-2014-CoG-646849Knut and Alice Wallenberg Foundation, 2012.0272Knut and Alice Wallenberg Foundation, 2015.0030Available from: 2019-11-28 Created: 2019-11-28 Last updated: 2019-11-28Bibliographically approved

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Alvarez, AlbertoMartinez-Corral, InesDaubel, NinaMäkinen, TaijaGängel, Konstantin

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