肢芽间充质干细胞条件性FKBP38基因敲除小鼠模型的构建

A preliminary study of conditional limb bud mesenchymal stem cell FKBP38 knockout mice

目的构建并鉴定条件性肢芽间充质干细胞FKBP38基因敲除小鼠,为研究FKBP38基因在骨发育及骨关节炎中的作用机制提供动物模型。方法将Prrx1-Cre小鼠与FKBP38^(flox/flox)交配繁殖出F1代小鼠,得到基因型为Prrx1-Cre(Fkbp38^(flox/+))小鼠;选基因型Prrx1-Cre(Fkbp38^(flox/+))的小鼠与FKBP38^(flox/flox)小鼠杂交获得F2代小鼠;基因型为Prrx1-Cre(Fkbp38^(flox/flox))的小鼠即为本实验所需要构建模型小鼠。3周龄鉴定小鼠基因型,4周龄时用Western blot验证FKBP38基因敲除效果。结果 F2代小鼠中包含基因敲除鼠,PCR结果显示基因型为Cre/FKBP38^(flox/flox),Western blot结果显示FKBP38基因敲除效果明显。结论基于Cre/loxp重组酶系统,繁殖出肢芽间充质干细胞FKBP38基因敲除鼠,为在动物水平上探究FKBP38在骨发育及骨关节炎的作用机制提供模型基础。

Objective To construct and identify conditional limb bud mesenchymal stem cell FKBP38 knockout mice,and provide an animal model for the mechanism of FKBP38 gene in bone development and osteoarthritis.Methods Prrx1-Cre mice were mated with FKBP^38flox/flox to produce F1 mice.The genotype was Prrx1-Cre(Fkbp^38flox/+)mice.The mice with genotype Prrx1-Cre(Fkbp^38flox/+)were selected.FKBP38^flox/flox mice were crossed to obtain F2 mice.Mice with genotype Prrx1-Cre(Fkbp^38flox/flox)were used to construct model mice for the experiment.The mouse genotype was identified at 3 weeks of age.The knockdown effect of FKBP38 gene was verified by Western blot at 4 weeks of age.Results F2 mice contained knockout mice.The PCR results showed that the genotype was Cre/FKBP^38flox/flox.The results of Western blot showed that the FKBP38 gene knockout effect was obvious.Conclusion Based on the Cre/loxp system,the limb bud mesenchymal stem cell FKBP38 knockout mouse is propagated to provide a model basis for exploring the mechanism of FKBP38 in bone development and osteoarthritis at the animal level.