应用双向启动子RNA干扰技术抑制ICOS表达  

Suppression of ICOS expression by bidirectional promoter RNA interference technology

作　　者：许雪青[1] 白云[1] 王丰[2] 王璞[1] 王艳艳[1] 熊加祥[1] 

机构地区：[1]第三军医大学基础医学部医学遗传学教研室 [2]第三军医大学西南医院检验科,重庆400038

出　　处：《免疫学杂志》2006年第1期13-16,共4页Immunological Journal

基　　金：国家自然科学基金资助项目(30300170;30271246)

摘　　要：目的应用双向启动子技术构建针对共刺激分子ICOS的逆转录病毒干扰载体,观察其对ICOS表达的影响。方法设计ICOS的干扰序列,构建基于双向启动子的逆转录病毒干扰载体MIW-ICOS,包装病毒后转染Hut78细胞,RT-PCR检测ICOS转录水平水平受抑制情况,FACS检测细胞表面ICOS蛋白表达受抑制情况。结果构建了针对ICOS三段序列的逆转录病毒干扰载体MIW-ICOS,并证实其中的一个干扰载体能有效抑制ICOS的表达。结论基于双向启动子的编号为245的ICOS逆转录病毒干扰载体能有效抑制Hut78 T细胞上ICOS的表达,为干预ICOS-ICOSL共刺激通路提供了有效手段。Objective To construct retrovirus vectors expressing siRNA for ICOS by bidirectional promoter RNA interference teclmology, and study its suppressing effect on ICOS. Methods Three sequence were designed and cloned into retrovirus vectors. After virus packaging, the T lymphocyte line Hut78 were transfected. The transcriptional level of ICOS were analyzed by RT-PCR and the ICOS expression level on cell surface were verified by FACS. Results Three RNA interference retrovirus vectors based on bidirectional promoter were constructed and one of them can suppress ICOS expression effectively. Conclusion The number 245 retrovirus vectors based on bidirectional promoter can suppress ICOS expression on Hut78 significantly, and the vector can be used as a tool for regulating ICOS-ICOSL pathway.

关 键 词：ICOS RNA干扰 双向启动子 

分 类 号：R392[医药卫生—免疫学]