Creation and characterization of BAC-transgenic mice with physiological overexpression of epitope-tagged RCAN1 (DSCR1).
Creation and characterization of BAC-transgenic mice with physiological overexpression of epitope-tagged RCAN1 (DSCR1).
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DOI：
10.1007/s00335-012-9436-9
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发表时间：
2013-02
期刊：
影响因子：
2.5
通讯作者：
Tycko, Benjamin
中科院分区：
文献类型：
--
作者：
Xing, Luzhou;Salas, Martha;Zhang, Hong;Gittler, Julia;Ludwig, Thomas;Lin, Chyuan-Sheng;Murty, Vundavalli V.;Silverman, Wayne;Arancio, Ottavio;Tycko, Benjamin
21号染色体基因RCAN 1编码钙调神经磷酸酶（CaN）的调节剂，是导致唐氏综合征（DS; 21三体）患者认知障碍的候选基因。为了开发用于研究RCAN 1的生物化学及其对DS的贡献的生理学相关模型，我们产生了含有人RCAN 1基因的细菌人工染色体转基因（BAC-Tg）小鼠系，该基因具有通过重组工程并入的C-末端HA-FLAG表位标签。BAC-Tg的表达水平仅略高于天然Rcan 1基因;在RCAN 1BAC-Tg 1中约为1.5倍，在RCAN 1BAC-Tg 2中约为2倍。从这些小鼠的脑中亲和纯化RCAN 1蛋白复合物，揭示了RCAN 1与钙调磷酸酶（CaN）、糖原合成酶激酶3-β（Gsk 3b）和钙调蛋白的核心复合物，其中亚化学计量组分包括LOC 73419。BAC-Tg小鼠是完全可行的，但在这些小鼠的海马脑切片中，长时程突触增强（LTP）与BAC-Tg剂量成比例地受损。RCAN 1在某些系统中可以作为肿瘤抑制因子，但我们发现RCAN 1 BAC-Tg在Tp 53;WAP-Cre小鼠中以低拷贝数存在时不会减少乳腺癌生长。这项工作建立了一个有用的小鼠模型，用于研究RCAN 1蛋白在体内的生物化学和剂量依赖性功能。
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 RCAN1BAC-Tg1 and 2-fold in RCAN1BAC-Tg2. Affinity purification of the RCAN1 protein complex from brains of these mice revealed a core complex of RCAN1 with calcineurin (CaN), glycogen synthase kinase 3-beta (Gsk3b), and calmodulin, with sub-stoichiometric components including LOC73419. The BAC-Tg mice are fully viable, but long-term synaptic potentiation (LTP) 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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DOI：
10.1093/jnen/63.5.429
发表时间：
2004-05-01
影响因子：
3.2
作者：
Branchi, I;Bichler, Z;Alleva, E
通讯作者：
Alleva, E
影响因子：
4.1
作者：
Genescà, L;Aubareda, A;Pérez-Riba, M
通讯作者：
Pérez-Riba, M
影响因子：
2.7
作者：
Lange, AW;Molkentin, JD;Yutzey, KE
通讯作者：
Yutzey, KE
影响因子：
4.8
作者：
Ermak, G;Morgan, TE;Davies, KJA
通讯作者：
Davies, KJA
影响因子：
30.8
作者：
LAMB, BT;SISODIA, SS;GEARHART, JD
通讯作者：
GEARHART, JD