title: [Construction  of recombinant lentiviral vector containing shRNA for human neuropilin-1 gene].
abstract: OBJECTIVE
  To construct and identify a recombinant lentiviral vector containing shRNA for human neuropilin-1 (NRP-1) gene.
  
  
  METHODS
  Four shRNA targeting the NRP-1 mRNA were designed to construct the pGCSIL-RFP-shNRP1 lentivirus vectors. The positive clone was chosen and confirmed by PCR and DNA sequencing. 293T cells were cotransfected with pGCSIL-RFP-shNRP1,  pHelper1.0 and pHelper 2.0 to package the lentivirus and the titer of the virus was tested. After lentivirus-shRNA and over-expression plasmid containing NRP-1 were transfected into 293T cells,  Western blotting was used to determine the expression of Flag gene in order to observe the inhibited efficacy of relative NRP-1 expression.
  
  
  RESULTS
  PCR analysis and DNA sequencing demonstrated that the shRNA sequence was consistent with the human NRP-1. The titer of the recombinant lentiviral vector was 1×10(9); Tu/mL. The relative expression of NRP-1 protein in the transfected cells significantly decreased after treated with lentiviral-shRNA.
  
  
  CONCLUSIONS
  We have constructed successfully the effective recombinant lentiviral vector containing shRNA for human NRP-1 gene.
authors.affiliation: Department of General Surgery, Xi'an Jiaotong University, Xi'an, China.
authors.forename: Jian-bao
authors.initials: JB
authors.lastname: Zheng
authors.forename: Zhi-min
authors.initials: ZM
authors.lastname: Geng
authors.forename: Qiang
authors.initials: Q
authors.lastname: Chen
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
mesh_terms: D001483:Base Sequence; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008969:Molecular Sequence Data; D039942:Neuropilin-1 / Q000235:genetics*; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D039942:Neuropilin-1