基于Cre/Loxp系统的Csf1r-CreERT2 R26REYFP报告基因小鼠的构建及效率检测
Construction and efficiency detection of Csf1r-CreERT2 R26REYFP reporter gene mouse based on Cre/Loxp system
朱向玲 1吴旭铭 1王卉卉 1周园园 1王安琪 1张慧茹 1刘崇 1涂佳杰1
作者信息
- 1. 安徽医科大学临床药理研究所,合肥 230032
- 折叠
摘要
目的 构建报告基因小鼠,评价Csf1r-CreERT2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-CreERT2小鼠与R26REYFP小鼠繁育,他莫昔芬诱导、PCR筛选Csf1r-CreERT2 R26REYFP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-CreERT2 R26REYFP报告基因小鼠.此外,Csf1r-CreERT2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R26REYFP组比较,Csf1 r-CreERT2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-CreERT2小鼠与R26REYFP小鼠是获得Csf1r-CreERT2 R26REYFP诱导型条件性荧光小鼠的有效途径.Csf1r-CreERT2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.
Abstract
Objective To construct Csf1r-CreERT2 R26REYFP reporter gene mice and assess the efficacy of Csf1r-CreERT2-mediated enhancement of CSF1R in CD45+cells labeled with yellow fluorescein protein EYFP.Methods Csf1r-CreERT2 mice were crossbred with R26REYFP homozygous mice,and Csf1r-CreERT2R26REYFP 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-CreERT2 R26REYFP reporter gene mice were acquired.In addition,it was found that Csf1r-CreERT2-mediated EYFP could effectively mark CSF1R in various tissues of mice and CD45+cells in different locations.Compared to the R26REYF P 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 sig-nificant difference was observed in the spleen tissue.Conclusion Adult Csf1r-CreERT2 mice and R26REYFP mice are effective ways to obtain Csf1r-CreERT2 R26REYFP induced conditional fluorescence mice.Csf1r-CreERT2 can mediate EYFP to effectively trace CSF1R in CD45+cells in different parts of mice.
关键词
Csf1r-CreERn/R26REYFP/Cre/LoxP/系统/CD45/流式细胞术Key words
Csf1r-CreERT2/R26REYFP/Cre/LoxP system/CD45/flow cytometry引用本文复制引用
基金项目
安徽省教育厅高校科学研究项目(2022AH020052)
出版年
2024
国家科技期刊平台
NSTL
万方数据