title: [Construction of VEGF gene-targeted small interfering RNA mediated by CMV promoter and its silencing effect].
abstract: To construct VEGF gene-targeted small interfering RNA (siRNA) and its expression vector driven by CMV promoter and to investigate its interference effect.
  
  
  
  The VEGF gene-targeted hairpin siRNA was designed, two complementary oligonucleotide strands were synthesized. After annealing, two-strand oligonucleotide was inserted into pDC311-SV40-RC vector, which was then identified by PCR and sequenced. Then human U-2 OS cell line was transfected with the vector using lipofectamine method. Finally, ELISA was performed to evaluate the expression of VEGF protein.
  
  
  
  PCR-identification of positive clone and sequencing confirmed the vector containing the target siRNA. ELISA showed that compared with the control group, the expression levels of VEGF protein in transfected U-2 OS cells were decreased significantly (P<0.05).
  
  
  
  VEGF gene-targeted siRNA and its vector mediated by CMV promoter were successfully constructed, which can reduce the VEGF protein expression after transfecting.
authors.affiliation: The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.
authors.forename: Xiao-yan
authors.initials: XY
authors.lastname: Liu
authors.forename: Hong
authors.initials: H
authors.lastname: Fang
authors.forename: Hong-chao
authors.initials: HC
authors.lastname: Chen
authors.forename: Xiao-ling
authors.initials: XL
authors.lastname: Jiang
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics; D001483:Base Sequence; D001859:Bone Neoplasms / Q000473:pathology; D045744:Cell Line, Tumor; D020868:Gene Silencing; D006801:Humans; D008969:Molecular Sequence Data; D012516:Osteosarcoma / Q000473:pathology; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A