Red2Flpe-SCON: a versatile, multicolor strategy for generating mosaic conditional knockout mice.
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Abstract
Image-based lineage tracing enables tissue turnover kinetics and lineage potentials of different adult cell populations to be investigated. Previously, we reported a genetic mouse model system, Red2Onco, which ectopically expressed mutated oncogenes together with red fluorescent proteins (RFP). This system enabled the expansion kinetics and neighboring effects of oncogenic clones to be dissected. We now report Red2Flpe-SCON: a mosaic knockout system that uses multicolor reporters to label both mutant and wild-type cells. We develop the Red2Flpe mouse line for red clone-specific Flpe expression, as well as the FRT-based SCON (Short Conditional IntrON) method to facilitate tunable conditional mosaic knockouts in mice. We use the Red2Flpe-SCON method to study Sox2 mutant clonal analysis in the esophageal epithelium of adult mice which reveal that the stem cell gene, Sox2, is less essential for adult stem cell maintenance itself, but rather for stem cell proliferation and differentiation.
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Acknowledgements: We thank present and past members of the Koo, Elling and Urban labs at IMBA for valuable discussions and critical comments, Dr. Rike Zietlow and the Life Science Editors for reading and correcting the manuscript, VBC core facilities (especially the Histopathology facility, BioOptics and the animal caretakers). We thank Single Cell Discoveries for helping us with single cell RNA sequencing and offering technical advice on sample handling. This work was supported by core funding from the Institute of Molecular Biotechnology (IMBA) of the Austrian Academy of Sciences; ERC starting grant, Troy Stem cells, 639050; Interpark Bio-Convergence Center Grant Program; and fellowship to S.W. (DOC Fellowship of the Austrian Academy of Sciences). G.C. was supported by a Lise Meitner Postdoctoral fellowship M2976, FWF, and by the FWF Standalone (P35694) and ERA PerMed (I 5900) grants. B.-K.K. and his team are supported by the Institute for Basic Science.
Funder: This work was supported by core funding from the Institute of Molecular Biotechnology (IMBA) of the Austrian Academy of Sciences; ERC starting grant, Troy Stem cells, 639050; Interpark Bio-Convergence Center Grant Program; and fellowship to first author Sam Wu (DOC Fellowship of the Austrian Academy of Sciences)
Funder: FWF Lise Meitner Postdoctoral fellowship M2976 FWF Standalone P35694 FWF ERA PerMed I 5900
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2041-1723
