title: [Construction and application of inducible artificial microRNA expression vector targeting eIF3g gene].
abstract: OBJECTIVE
  To construct an inducible artificial microRNA expression vector targeting eIF3g gene and use it to inhibit the expression of eIF3g in K562 cells.
  
  
  METHODS
  The microRNA targeting human eIF3g was designed and obtained by PCR. After confirmed by sequencing, the microRNA was cloned into the pRevTRE2 plasmid. The Tet-off plasmids were transfected into K562 cells and selected for the stable tetracycline inducible K562/Tet-off transfectants. The pRevTRE2-eIF3g-miRNA plasmid was then transfected into stable K562/Tet-off transfectants and the expression of eIF3g was detected by Western blot.
  
  
  RESULTS
  The microRNA targeting eIF3g was confirmed by sequencing. GFP fluorescence assay showed that the stable transfectants of Tet-off plasmids were under tetracycline control. Western blot results showed that the stable transfectants of pRevTRE2-eIF3g-microRNA inhibited eIF3g expression under the regulation of tetracycline.
  
  
  CONCLUSIONS
  The tetracycline-inducible artificial microRNA expression vector targeting eIF3g gene has been successfully constructed and effectively inhibits eIF3g expression in K562 cells.
authors.affiliation: Department of General Surgery, Sir Run Run Shaw Hospital, Zhejiang University and Key Laboratory of Biotherapy of Zhejiang Province, Hangzhou 310016, China. 2008chulisha@163.com
authors.forename: Li-Sha
authors.initials: LS
authors.lastname: Chu
authors.forename: Na
authors.initials: N
authors.lastname: Han
authors.forename: Xiang-Yang
authors.initials: XY
authors.lastname: Song
authors.forename: Jiang
authors.initials: J
authors.lastname: Cao
mesh_terms: D039621:Eukaryotic Initiation Factor-3 / Q000235:genetics* / Q000378:metabolism*; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D035683:MicroRNAs / Q000235:genetics*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013752:Tetracycline / Q000494:pharmacology
chemical_list: D039621:Eukaryotic Initiation Factor-3; D035683:MicroRNAs; D011500:Protein Synthesis Inhibitors; D013752:Tetracycline