title: [Construction and evaluation of human Dp71 shRNA vector].
abstract: OBJECTIVE
  To construct effective short hairpin RNA (shRNA) recombinant plasmids targeting human Dystrophin Dp71 gene, and evaluate their interference efficiency.
  
  
  METHODS
  Three pairs of siRNA sequences targeting human Dp71 gene and one pair of control siRNA sequence were designed, synthesized, and then inserted into the pRNAT-U6.1/Neo vector. The shRNA recombinant vectors were evaluated by enzyme digestion and sequencing. Dp71-shRNA and control shRNA plasmids were transfected into human normal gastric epithelial cells (GES-1) and human bronchial epithelium (HBE). Western blot was used to evaluate its interfering efficiency.
  
  
  RESULTS
  Restriction enzyme digestion and sequencing showed that the Dp71-shRNA vectors were successfully constructed. Western blot displayed that Dp71 protein expression was reduced to a significant degree after transfection with the 3 Dp71-shRNA plasmids, and Dp71-shRNA2 plasmid inhibit the Dp71 expression most efficiently.
  
  
  CONCLUSIONS
  Dp71-shRNA vectors have been successfully constructed. The 3 Dp71-shRNA plasmids can inhibit Dp71 expression in GES-1 and HBEC, with Dp71-shRNA2 plasmid displaying the highest inhibition efficiency.
authors.affiliation: Department of Cardiovascular and Thoracic Surgery, Second Xiangya Hospital, Central South University, Changsha 410011,China.
authors.forename: Sichuang
authors.initials: S
authors.lastname: Tan
authors.forename: Zhikang
authors.initials: Z
authors.lastname: Chen
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Cao
authors.forename: Qiaocheng
authors.initials: Q
authors.lastname: Wen
authors.forename: Weilin
authors.initials: W
authors.lastname: Zhang
authors.forename: Qingren
authors.initials: Q
authors.lastname: Zeng
authors.forename: Sipin
authors.initials: S
authors.lastname: Tan
mesh_terms: D016189:Dystrophin / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
chemical_list: D016189:Dystrophin; D034741:RNA, Small Interfering; C085339:apo-dystrophin 1