中国免疫学杂志Issue(2)：151-155,5.DOI:10.3969/j.issn.1000-484X.2015.02.002
Mcl-1短发夹RNA在Raw264.7细胞内特异性的筛选
Specificity of screening of short hairpin RNA targeting Mcl-1 gene in Raw264.7 cells
摘要
Abstract
Objective:To transfect Mcl-1shRNA into Raw264.7 cells,and screen out specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene to figure out the effect of shRNA on Mcl-1 expression in murine macrophage cell line Raw 264.7.Methods: Specific shRNA was transfected into murine macrophage cell line Raw 264.7 via lipofectamine.Semi-quantitative RT-PCR and Western blot were respectively employed to test the changes in Mcl-1 mRNA level and Mcl-1 protein expressions 24 h and 48 h after the transfection ,and the silencing effects of the three pairs of specific shRNA fragments corresponding to different sites were analyzed.Results: Specific shRNA fragments at 24 h and 48 h could effectively reduce Mcl-1 mRNA and protein level ,with higher silencing effects than those of the normal group ,the lipofectamine group and the negative control group.There were statistically significant differences among them ( P<0.05 ).Among the three pairs of specific shRNA fragments corre-sponding to different sites ,Mcl-1 shRNA3 showed the most significant inhibiting effect on Mcl-1 mRNA and proteins.Conclusion:RNA interference can downregulate the level of Mcl-1 mRNA in murine macrophage cell line Raw 264.7 and greatly downregulate the expression of Mcl-1protein.Specific shRNA eukaryotic expression plasmids with the most significant effect of silent Mcl-1 gene have been screened out successfully.关键词
Mcl-1/RNA干扰/Raw264.7Key words
Mcl-1/RNA interference/Raw264.7分类
医药卫生引用本文复制引用
王婵,王新敏,王飞雨,张雨晴,曹旭东,吴江东,吴芳,张万江,章乐..Mcl-1短发夹RNA在Raw264.7细胞内特异性的筛选[J].中国免疫学杂志,2015,(2):151-155,5.基金项目
本文为国家自然科学基金资助项目（No.81260241）和石河子大学项目（RCZX200922）。 （）
