title: [shRNA mediated silencing effect of RNA interference on the CD28 co-stimulation factor in mice T lymphocyte].
abstract: OBJECTIVE
  To explore the silencing effect of short hairpin RNA (shRNA) on the CD28 of mice T lymphocytes by CD28-shRNA expressing plasmid evaluate the interfering effects (chronology and stability) mediated by shRNA and select out the most efficient CD28 shRNA sequence.
  
  
  METHODS
  Three CD28 specific and one non-specific shRNA expressing plasmids were constructed and then transfected separately into mice spleen T lymphocytes. Non-transfected cells and non-specific shRNA were taken as controls. Inhibitory effect of CD28 shRNA was demonstrated by real-time quantitative PCR and Western blots. The sequence of the highest RNA interference (RNAi) efficacy was screened.
  
  
  RESULTS
  (1) CD28 shRNA expressing plasmids were successfully constructed; (2) Three CD28 specific shRNAs effectively inhibited the expression of CD28 at the mRNA and protein levels, and there was a statistically significant difference comparing with the controls (P < 0.01): The copies of CD28 in mice spleen cells at the mRNA levels were persistently decreased by 99.62%, 99.89% and 99.80% respectively after 20 days, and so did at the protein level [(84.90 +/- 0.65)%, (96.49 +/- 0.03)%, (91.76 +/- 0.32)% respectively]. The highest inhibitory rate was in CD28 shRNA-2 group.
  
  
  CONCLUSIONS
  (1) Specific shRNA can mediate long-term and stable silencing effects on CD28 gene; (2) shRNAs matching different sites of CD28 gene exert differential inhibitory effects.
authors.affiliation: Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Sang
authors.forename: Kai-lin
authors.initials: KL
authors.lastname: Xu
authors.forename: Xiu-ying
authors.initials: XY
authors.lastname: Pan
authors.forename: Ling-yu
authors.initials: LY
authors.lastname: Zeng
authors.forename: Bing
authors.initials: B
authors.lastname: Du
mesh_terms: D000818:Animals; D018106:CD28 Antigens / Q000235:genetics*; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D013601:T-Lymphocytes / Q000378:metabolism*; D014162:Transfection
chemical_list: D018106:CD28 Antigens