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Creation and characterization of BAC-transgenic mice with physiological overexpression of epitope-tagged RCAN1 (DSCR1)

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Abstract

The chromosome 21 gene RCAN1, encoding a modulator of the calcineurin (CaN) phosphatase, is a candidate gene for contributing to cognitive disability in people with Down syndrome (DS; trisomy 21). To develop a physiologically relevant model for studying the biochemistry of RCAN1 and its contribution to DS, we generated bacterial artificial chromosome-transgenic (BAC-Tg) mouse lines containing the human RCAN1 gene with a C-terminal HA-FLAG epitope tag incorporated by recombineering. The BAC-Tg was expressed at levels only moderately higher than the native Rcan1 gene: approximately 1.5-fold in RCAN1 BAC-Tg1 and twofold in RCAN1 BAC-Tg2. Affinity purification of the RCAN1 protein complex from brains of these mice revealed a core complex of RCAN1 with CaN, glycogen synthase kinase 3-beta (Gsk3b), and calmodulin, with substoichiometric components, including LOC73419. The BAC-Tg mice are fully viable, but long-term synaptic potentiation is impaired in proportion to BAC-Tg dosage in hippocampal brain slices from these mice. RCAN1 can act as a tumor suppressor in some systems, but we found that the RCAN1 BAC-Tg did not reduce mammary cancer growth when present at a low copy number in Tp53;WAP-Cre mice. This work establishes a useful mouse model for investigating the biochemistry and dose-dependent functions of the RCAN1 protein in vivo.

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Acknowledgments

This work was supported by grants from the NIH (P01-HD035897 to BT and WS, and R01-AG034248 to OA).

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  1. Thomas Ludwig

    Present address: Department of Molecular & Cellular Biochemistry, Ohio State University Medical Center, Columbus, OH, USA

Authors and Affiliations

  1. Institute for Cancer Genetics, Columbia University Medical Center, Herbert Irving Cancer Research Building, Room 607, New York, NY, 10032, USA

    Luzhou Xing, Martha Salas, Julia Gittler, Thomas Ludwig, Vundavalli V. Murty & Benjamin Tycko

  2. Department of Pathology, Columbia University Medical Center, New York, NY, 10032, USA

    Hong Zhang, Vundavalli V. Murty, Ottavio Arancio & Benjamin Tycko

  3. Herbert Irving Comprehensive Cancer Center, Columbia University Medical Center, New York, NY, 10032, USA

    Chyuan-Sheng Lin, Vundavalli V. Murty & Benjamin Tycko

  4. Kennedy-Krieger Institute, Baltimore, MD, 21202, USA

    Wayne Silverman

  5. Taub Institute for Research on Alzheimer’s Disease and the Aging Brain, Columbia University Medical Center, New York, NY, 10032, USA

    Ottavio Arancio & Benjamin Tycko

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  1. Luzhou Xing
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  2. Martha Salas
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  3. Hong Zhang
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Correspondence to Benjamin Tycko.

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Xing, L., Salas, M., Zhang, H. et al. Creation and characterization of BAC-transgenic mice with physiological overexpression of epitope-tagged RCAN1 (DSCR1). Mamm Genome 24, 30–43 (2013). https://doi.org/10.1007/s00335-012-9436-9

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