title: [Construction of beta-catenin miRNA-expressing vectors and test of silencing effect in adipose-derived stem cells from Sprague-Dawley rats].
abstract: OBEJECTIVE: To construct miRNA-expressing plasmid vector for interfering the expression of beta-catenin in adipose-derived stem cells from Sprague-Dawley rats.
  
  
  METHODS
  The double strands oligonucleotides of miR-expressing cassette were ligated into plasmid pSWH to generate pSWH-miR. Then the PCR product of enhanced green fluorescence protein (EGFP) open reading frame (ORF) fragment was inserted into BamH I and Xbo I digested pSWH-miR to generate pSWH-EGFP-miR. The adipose-derived stemcells from rats (rADSCs) were transfected with pSWH-EGFP-miR respectively. The expression of beta-catenin was determined by Western blot at 72 h post-transfection.
  
  
  RESULTS
  Five miRNA-expressing plasmid vectors were constructed (miR-780, miR-796, miR-1467, miR-1948, miR-1960). miR-780 and miR-796 had the best silencing effect on the expression of beta-catenin (P < 0.05), less did the miR-1960 (P < 0.05), but not the miR-1467 and the miR-1948 (P < 0.05).
  
  
  CONCLUSIONS
  miR-780 and miR-796 could interference the beta-catenin in ADSCs with high efficiency.
authors.forename: Hai-Sen
authors.initials: HS
authors.lastname: Huang
authors.forename: Shuang
authors.initials: S
authors.lastname: Liu
authors.forename: Mei
authors.initials: M
authors.lastname: Yu
authors.forename: Xiao-Xin
authors.initials: XX
authors.lastname: Luo
authors.forename: Wei-Hua
authors.initials: WH
authors.lastname: Guo
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Tian
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D020868:Gene Silencing; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000166:cytology*; D014162:Transfection; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism*
chemical_list: D035683:MicroRNAs; D051176:beta Catenin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins