摘要
目的利用FLP/FRT、Cre/Loxp重组酶系统构建并鉴定髓样细胞特异性SETD4基因敲除小鼠,为深入研究SETD4的生物学功能奠定基础。方法将引进的Setd4^(flox/+)小鼠自交,筛选出子代基因型为Setd4^(flox/flox)的小鼠;与FLP小鼠交配,得到Setd4^(fl/+/flp)小鼠;然后分别与C57BL/6小鼠交配去除FLP酶,筛选出Setd4^(fl/+)小鼠;与Lyz2-Cre小鼠交配,筛选出Setd4^(fl/+)/Lyz2-Cre小鼠;将得到的Setd^(4fl/+)和Setd4^(fl/+)/Lyz2-Cre小鼠交配,筛选出Setd4^(-/-)/Lyz2-Cre小鼠,即髓样细胞特异性SETD4基因敲除小鼠。利用PCR技术鉴定小鼠基因型;实时荧光定量PCR技术检测小鼠腹腔巨噬细胞及肝组织中SETD4的mRNA表达水平验证敲除情况。结果髓样细胞特异性SETD4基因敲除小鼠腹腔巨噬细胞中SETD4 mRNA水平较野生型小鼠显著降低;而在肝组织中无显著差异。结论利用FLP/FRT、Cre/Loxp系统成功构建髓样细胞特异性SETD4基因敲除小鼠,为后续的功能学研究提供了动物模型。
Objective To elucidate the function of SETD4, we established myeloid cell-specific SETD4 knockout mice. Methods Setd4^flox/＋ mice were inbred to obtain Setd4^flox/flox mice, which then bred with FLP mice to obtain the Setd4^fl/＋/flp mice. After crossing with C57BL/6 mice, Setd4^fl/＋ mice were obtained,while Setd4^fl/＋/Lyz2-Cre were obtained after crossing with Lyz2-Cre mice. And then Setd4^-/- /Lyz2-Cre mice were generated by mating Setd4^fl/＋ /Lyz2-Cre with Setd4^fl/＋ mice. PCR was used to identify the genotype of offspring. The mRNA level of SETD4 in peritoneal macrophages and liver were detected by quantitative real time PCR to confirm the knockout efficiency.Result The mRNA expression of SETD4 in peritoneal macrophages of myeloid cell-specific SETD4 knockout mice was significantly lower than the wildtype mice,while no significant difference could be detected in the liver. Conclusion Based on FLP/FRT、Cre/Loxp recombination system, myeloid cell-specific SETD4 knockout mice were successfully established for further research.
出处
《中国临床解剖学杂志》
CSCD
北大核心
2017年第2期177-182,共6页
Chinese Journal of Clinical Anatomy
基金
国家自然科学基金项目(81471901
81072425)
广东省自然科学基金重点项目(2015A030311031)
