title: [Construction of RNA interference expression vectors of human neuropathy target esterase and its inhibition for expression of NTE in mammalian cells].
abstract: OBJECTIVE
  To construct the RNA interference expression vector for expression of human neuropathy target esterase (NTE) gene in mammalian cells.
  
  
  METHODS
  Spe I and Xho I-digested insert from pSUPER, which comprised H1 RNA polymerase III promoter and the multiple cloning sites, were cloned into the compatible in the pcDNA3.1 (+) to generate pSUPER/neo that could express small interfering RNA in mammalian cells. The annealed oligos targeting the expression of NTE were ligated into pSUPER/neo vector digested with Bgl II and Hind III to generate pSUPER/neo-NTE, which was transfected into COS7 and SH-SY5Y cells. The inhibitory effect of the expression of NTE was detected by western blot analysis and the enzyme activity assay.
  
  
  RESULTS
  pSUPER/neo-NTE could stably express double-stranded RNA of NTE. The expression of pSUPER/neo-NTE in COS7 and SH-SY5Y cells could efficiently inhibit the activity of NTE in the mammalian cells.
  
  
  CONCLUSIONS
  Stable eukaryotic expression vector of double-stranded RNA of NTE, pSUPER/neo-NTE, has been constructed successfully with promoter substitution strategy.
authors.affiliation: Laboratory of Molecular Toxicology, State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China.
authors.forename: Ping-an
authors.initials: PA
authors.lastname: Chang
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Yi-jun
authors.initials: YJ
authors.lastname: Wu
mesh_terms: D000818:Animals; D019556:COS Cells; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D005822:Genetic Vectors; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering; D002265:Carboxylic Ester Hydrolases; C021859:neurotoxic esterase