山西大学学报（自然科学版）2012，Vol.35Issue(2)：376-382,7.
重组慢病毒介导的NMD途径因子UPF1和SMG1可诱导干扰细胞株的构建
Construction of Cell Strain Containing Inducible Interference of NMD Factor UPF1 and SMG1 Mediated by Lentiviral System
摘要
Abstract
Nonsense-mediated decay (NMD) is well known by the lucid definition of being a RNA surveillance mechanism that ensures the speedy degradation of mRNAs containing premature translation termination codons. The definition of aberrant transcription and start of NMD are determined by the protein complex binding on EJC,where UPF1 and SMG1 play key role for NMD path. UPF1 is a RNA helicase and a RNA-dependent ATPase. SMG1 is responsible for UPF1 phosphorylation as a phosphatidylinositol 3-ki-nase. In this study,we construct the pHIV-7-derived lentiviral vector pTIG (pHIV7-TetR-IRES-GFP) encoding a U6tetOpromoted short hairpin RNA (shRNA) cassette containing UPF1 and SMG1. The resultant plasmids were transfected into HEK293T cell with the help of packaging plasmids to result in recombi-nant virus harboring hairpin RNA cassette. The resultant virus were then used to infect AD_293 cell to observe the expression of hairpin RNA gene. We have obtained the cell strains expressing hairpin RNA of UPF1 and SMGl,and these cell strains could be used for next research on the mechanism of NMD and gene screen.关键词
NMD途径/UPF1/SMG1/PTC/RNA干扰/shRNAKey words
NMD path/UPF1/SMG1/ premature termination codon/RNA interference/shRNA分类
生物科学引用本文复制引用
柴宝峰,常文娟,申泉,王刚..重组慢病毒介导的NMD途径因子UPF1和SMG1可诱导干扰细胞株的构建[J].山西大学学报（自然科学版）,2012,35(2):376-382,7.基金项目
国家自然科学基金(31172078 （）
30770294) （）
山西省自然科学基金(2009011040-1) （2009011040-1）
山西省留学归国基金项目 （）
