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Generation and characterization of a complete null estrogen receptor α mouse using Cre/LoxP technology

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Abstract

Conventional estrogen receptor α knockout (neo-ERαKO, neo-ERα−/−) mice contain a truncated and chimeric ERα fusion protein that retains 35% estrogen-dependent transactivation activity, and therefore the in vivo ERα function is difficult to study thoroughly. Furthermore, these neo-ERα−/− mice cannot be used for tissue and temporal specific ERα deletion. Therefore, there is a clear need to establish a floxed ERα mouse line that can knockout ERα specifically and completely in each selected cell type. Here we generated floxed ERα mice using a self-excising ACN (tACE-Cre/Neo) cassette. Mating the floxed ERα mice with ACTB-Cre mice produces a deletion of the floxed allele disrupting the reading frame of the ERα transcript so that no ERα protein is detected in the ACTB-Cre/ERα−/− mice. Expression of ERα target genes, such as G-6-PD and lactoferrin, is diminished by over 90% in the ACTB-Cre/ERα−/− uterus, but not in the neo-ERα−/− uterus. Furthermore, we also validated that ACTB-Cre/ERα−/− females have a hypoplastic internal genital tract, polycystic ovaries with hemorrhagic follicles, infertility, and higher body weight. Together, our data clearly demonstrate that the newly established floxed ERα mouse is a reliable mouse model for future studies of ERα roles in vivo in the selective estrogen target tissues. The complete knockout of ERα in the ACTB-Cre/ERα−/− mice will also provide an improved mouse model to study the role of ERα in vivo.

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Acknowledgments

This work was partly supported by NIH grant DK60912. This research was also supported in part by NICHD/NIH through cooperative agreement (U54 HD 933067, the Baltimore-Chicago Center for Reproductive Research) as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research (SCCPIR). The technical assistance of Yayoi Shibusawa is gratefully acknowledged. We also thank Karen Wolf and Susan R. Schoen for assisting in manuscript preparation.

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Authors and Affiliations

  1. George Whipple Lab for Cancer Research, Departments of Pathology and Urology, University of Rochester Medical Center, 601 Elmwood Ave, Box 656, Rochester, NY, 14642, USA

    Ming Chen, Xi Wang, Chawnshang Chang & Shuyuan Yeh

  2. Department of Pediatrics, Johns Hopkins University School of Medicine, 600 North Wolfe Street, CMSC 406, Baltimore, MD, 21287, USA

    Andrew Wolfe & Sally Radovick

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  1. Ming Chen
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  2. Andrew Wolfe
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  3. Xi Wang
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  4. Chawnshang Chang
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  5. Shuyuan Yeh
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Correspondence to Shuyuan Yeh or Sally Radovick.

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M. Chen and A. Wolfe contributed equally to this article.

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Chen, M., Wolfe, A., Wang, X. et al. Generation and characterization of a complete null estrogen receptor α mouse using Cre/LoxP technology. Mol Cell Biochem 321, 145–153 (2009). https://doi.org/10.1007/s11010-008-9928-9

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  • DOI: https://doi.org/10.1007/s11010-008-9928-9

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