首页|基于Cre/Loxp系统的Csf1r-CreERT2 R26REYFP报告基因小鼠的构建及效率检测

基于Cre/Loxp系统的Csf1r-CreERT2 R26REYFP报告基因小鼠的构建及效率检测

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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。
Construction and efficiency detection of Csf1r-CreERT2 R26REYFP reporter gene mouse based on Cre/Loxp system
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.

Csf1r-CreERT2R26REYFPCre/LoxP systemCD45flow cytometry

朱向玲、吴旭铭、王卉卉、周园园、王安琪、张慧茹、刘崇、涂佳杰

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安徽医科大学临床药理研究所,合肥 230032

Csf1r-CreERn R26REYFP Cre/LoxP 系统 CD45 流式细胞术

安徽省教育厅高校科学研究项目

2022AH020052

2024

安徽医科大学学报
安徽医科大学

安徽医科大学学报

CSTPCD北大核心
影响因子:1.095
ISSN:1000-1492
年,卷(期):2024.59(7)
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