利用Sleeping beauty转座子系统构建响应IFNβ的荧光转基因小鼠  

Effective Generation of the Fluorescent Mouse Model Responding to IFNβ Signal using the Sleeping Beauty Transposon System

作　　者：蔡少丽[1] 许姗[1] 焦长峰 丰志华 梁旺旺 邓湘凌 陈骐[1] 沈阳坤[1] CAI Shaoli;XU Shan;JIAO Changfeng;FENG Zhihua;LIANG Wangwang;DENG Xiangling;CHEN Qi;SHEN Yangkun(Fujian Key Laboratory of Innate Immune Biology,Biomedical Research Center of South China,Fujian Normal University,Fuzhou 350117,China)

机构地区：[1]福建师范大学南方生物医学研究中心,福建省天然免疫生物学重点实验室,福州350117

出　　处：《中国细胞生物学学报》2022年第9期1763-1771,共9页Chinese Journal of Cell Biology

基　　金：福建省自然科学基金(批准号:2017J01621、2021J01202)资助的课题。

摘　　要：干扰素β(interferon β,IFNβ)在抗病毒感染、免疫调节及肿瘤治疗中发挥重要作用。近年来,越来越多的证据表明IFNβ能显著增强树突状细胞(dendritic cell,DC)的抗原呈递能力以及T细胞的活性。然而目前针对IFNβ在病毒入侵和肿瘤发生过程中如何诱发全身性免疫应答的机制尚不清楚,这极大地限制了大家对病毒感染和肿瘤免疫的理解。因此,在这一过程中检测释放IFNβ的细胞种类以及IFNβ在各组织中的表达水平显得尤为重要。目前将外源DNA序列插入到宿主基因组中仍然需要合成或克隆同源模板,这会花费大量的时间成本,且效率不高。在这里,该研究利用Sleeping beauty转座子/转座酶系统构建了由IFNβ启动子诱导表达红色荧光蛋白的转基因小鼠模型(C57BL/6)。通过PCR及流式细胞术证实,小鼠中的外源基因经过数次传代仍然稳定表达。这些结果表明,该研究成功构建了由IFNβ启动子诱导稳定表达mCherry的转基因小鼠模型。这一模型为进一步研究干扰素通路在抗肿瘤和抗病毒中的功能提供有用的工具。IFNβ (interferon β) has an indispensable role in antiviral,immune regulation and oncological therapy.In recent years,numerous evidences have suggested that IFNβ can improve the capacity of the antigen presentation of DC (dendritic cell),as well as significantly increase the activity of T cells.However,it is not clear how IFNβ induces the systematic immune responses during viral infection and tumorigenesis.For this reason,it is vital to find out what types of cells release IFNβ and detect the expression level of IFNβ in various organs.Here,the “Sleeping beauty” transposon/transposase system is applied to construct a genetically modified mouse that is able to express mCherry driven by the IFNβ promoter.The exogenous sequences remain unchanged and expressed following several generation in the mice as verified by PCR and flow cytometry.These data indicate that the mouse model enabling stable expression of mCherry controlled by the IFNβ promoter is successfully constructed.This mouse model provides a useful tool for studying the regulation and function of IFNβ signaling pathway related to its antiviral and antitumor activities.

关 键 词：干扰素Β Sleeping beauty转座子系统 MCHERRY 转基因小鼠 

分 类 号：R730.51[医药卫生—肿瘤]