基于Cre/Loxp系统的Csf1r-Cre^(ERT2)R26R^(EYFP)报告基因小鼠的构建及效率检测

Construction and efficiency detection of Csf1r-Cre^(ERT2)R26R^(EYFP)reporter gene mouse based on Cre/Loxp system

目的构建报告基因小鼠,评价Csf1r-Cre^(ERT2)介导增强黄色荧光素蛋白EYFP标记组织CD45^(+)细胞CSF1R的效率。方法Csf1r-Cre^(ERT2)小鼠与R26R^(EYFP)小鼠繁育,他莫昔芬诱导、PCR筛选Csf1r-Cre^(ERT2)R26R^(EYFP)小鼠,流式细胞术和Western blot分析EYFP对不同组织以及不同组织CD45^(+)细胞中CSF1R的标记效率。结果获得Csf1r-Cre^(ERT2)R26R^(EYFP)报告基因小鼠。此外,Csf1r-Cre^(ERT2)小鼠介导EYFP可有效标记小鼠组织CSF1R以及不同部位中CD45^(+)细胞。与R26R^(EYFP)组比较,Csf1r-Cre^(ERT2)小鼠介导EYFP标记效率最高的是脑组织(P<0.001),最低的是胸腺组织(P<0.05),脾脏组织则差异无统计学意义。结论成年Csf1r-Cre^(ERT2)小鼠与R26R^(EYFP)小鼠是获得Csf1r-Cre^(ERT2)R26R^(EYFP)诱导型条件性荧光小鼠的有效途径。Csf1r-Cre^(ERT2)介导EYFP可对小鼠不同部位CSF1R以及CD45^(+)细胞中CSF1R进行有效示踪。

Objective To construct Csf1r-Cre^(ERT2)R26R^(EYFP)reporter gene mice and assess the efficacy of Csf1r-Cre^(ERT2)-mediated enhancement of CSF1R in CD45^(+)cells labeled with yellow fluorescein protein EYFP.Methods Csf1r-Cre^(ERT2)mice were crossbred with R26R^(EYFP)homozygous mice,and Csf1r-Cre^(ERT2)R26R^(EYFP)mice were identified through PCR and Western Blot analyses.Flow cytometry was employed to evaluate CSF1R tag-efficiency in CD45^(+)cells across different mouse tissues following tamoxifen induction.Results Csf1r-Cre^(ERT2)R26R^(EYFP)reporter gene mice were acquired.In addition,it was found that Csf1r-Cre^(ERT2)-mediated EYFP could effectively mark CSF1R in various tissues of mice and CD45^(+)cells in different locations.Compared to the R26R^(EYFP)group,the highest labeling efficiency was observed in the brain tissue(P<0.001),the lowest in the thymus tissue(P<0.05),and no significant difference was observed in the spleen tissue.Conclusion Adult Csf1r-Cre^(ERT2)mice and R26R^(EYFP)mice are effective ways to obtain Csf1r-Cre^(ERT2)R26R^(EYFP)induced conditional fluorescence mice.Csf1r-Cre^(ERT2)can mediate EYFP to effectively trace CSF1R in CD45^(+)cells in different parts of mice.