军事医学2012，Vol.36Issue(11)：825-829,5.
人THAP11基因RNAi慢病毒表达系统的建立
Construction of lentiviral vectors containing THAP11 RNAi and establishment of expression systems
摘要
Abstract
Objective To prepare the lentivirus granules interfering human THAP11 gene and knock down the expression of endogenous THAP11 in K562 cells and primary cord blood CD34+ cells. Methods DNA segments carrying the specific THAP11 interfering sequences were cloned into the plasmid pSicoR to generate siTHAPl 1 -pSicoR vectors. The siT-HAP11-pSicoR plasmids and three packing plasmids pLP1, pLP2 and pLP/VSVG were transfected into 293 T cells to produce lentivirus granules interfering THAP11. The siTHAPl 1-K562 cell line was established after infecting the lentivirus granules interfering THAP11 and the interference effect was evaluated by Western blot. Primary cord blood CD34+ cells were infected with lentivirus granules and the expression of THAP11 was investigated by real-time PCR. Results Two lentivirus vectors carrying the interfering sequence against THAP11 were successfully generated and the titer of the lentivirus was 1. 8 ×108 TU/ml. In K562 cells, THAP11 protein and mRNA levels were knocked down by the two siRNA lentivirus vectors. In CD34+ cells infected with siTHAPl 1-1 and siTHAPl 1-2, THAP11 mRNA levels were downregulated by 80% and 85% , respectively. Conclusion Human THAP11 siRNA lentiviral granules have been successfully generated.关键词
THAP11/RNA干扰/慢病毒/细胞系/人造血干细胞Key words
THAP11/ RNAi/ lentivirus/ cell line/ HSC分类
医药卫生引用本文复制引用
孔祥祯,尹荣华,郑巍薇,詹轶群,杨晓明,李长燕..人THAP11基因RNAi慢病毒表达系统的建立[J].军事医学,2012,36(11):825-829,5.基金项目
国家973计划资助项目(2010CB911904),国家863计划资助项目(2012AA020206) （2010CB911904）
