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| 慢病毒载体介导的RNA干扰抑制PC12细胞MyD88基因的表达 | |
| 引用本文： | 崔万鹏,刘晓湘,方秀斌. 慢病毒载体介导的RNA干扰抑制PC12细胞MyD88基因的表达[J]. 解剖科学进展, 2010, 16(2): 131-134 |
| 作者姓名： | 崔万鹏 刘晓湘 方秀斌 |
| 作者单位： | 中国医科大学,基础医学院神经生物学教研室,辽宁,沈阳,110001 |
| 基金项目： | 辽宁省高等学校科研项目(No.2008813) |
| 摘 要： | 目的研究慢病毒介导的RNA干扰对PC12细胞的转染效率和对MyD88基因的抑制效率。方法构建针对大鼠MyD88基因3个靶点的ShRNA慢病毒载体,PC12细胞按照不同的转染条件分为正常组(DMEM完全培养液)、DMEM加入polybrene组、EN.iS(Enhance infection solution)组、EN.iS加入polybrene组。荧光显微镜下观察不同组的转染效率,采用Real-timePCR和Western blot检测不同的靶点对MyD88的不同沉默效率。结果病8毒滴度为1×10TU/ml,MOI为100时,PC12细胞在EN.iS组的转染效率最高,转染试剂polybrene对转染效率无明显影响,沉默效率最高的靶点是5'-CATAC GCAACCAGCAGAAA-3'。结论慢病毒介导的RNA干扰是一种高效的基因沉默手段,可以对PC12细胞中MyD88的功能进行长期的研究。 |
| 关 键 词： | PC12细胞 慢病毒 RNA干扰 髓样分化因子初次应答基因88 |
Lentivirus mediated RNA interference knockdowns the expression of MyD88 in PC12 cells |
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| CUI Wan-peng,LIU Xiao-xiang,FANG Xiu-bin. Lentivirus mediated RNA interference knockdowns the expression of MyD88 in PC12 cells[J]. Progress of Anatomical Sciences, 2010, 16(2): 131-134 | |
| Authors: | CUI Wan-peng LIU Xiao-xiang FANG Xiu-bin |
| Affiliation: | Department of Neurobiology;China Medical University;Liaoning Shenyang 110001 China |
| Abstract: | Objective To investigate the transfection efficiency of lentivirus and silencing efficiency of MyD88 by lentivirus mediated RNA interference in PC12 cell line.Methods Recombinant lentivirus vectors with shRNA targeting rat myeloid differentiation primary response gene 88(MyD88)were constructed and transfected into pheochromocytoma cells(PC12)in vitro,the tranfection efficiency was observed under fluorescence microscope and the expression of MyD88 was 8 determined by Real-time PCR and Western Blot respective... |
| Keywords: | Pc12 cell line lentivirus RNA interference myeloid differentiation primary response gene88 |
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