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Objective: RNAi (RNA interference) technology of targeting α1, 3 galactosyltransferase enzyme (α1, 3GT) of shRNA lentiviral vectors inhibit vascular endothelial cells α1, 3GT and α1, 3 galactose residues Galα ( 1,3) Gal expression feasible. Methods: Using tissue explant cultured mouse endothelial cells; designed and synthesized by in vitro against α1, 3GTmRNA sequence-specific shRNA, and build portable α1, 3GT shRNA recombinant lentiviral vector, a lentivirus infected mice hemangioma endothelial cell line EOMA; fluorescence real-time quantitative PCR after transfection α1, 3GT mRNA expression level, immunofluorescence xenoantigenic Galα (1,3) Gal expression level; using SPSS 13.0 for Windows for data analysis. Results: In vitro tissue explant culture obtained high purity mouse vascular endothelial cells; successfully constructed recombinant lentiviral vector plasmid. The virus package, has been carrying α1, 3GT-shRNA mature recombinant lentiviral particles; fluorescence real-time quantitative PCR showed that recombinant lentiviral transfection EOMA, can inhibit the α1, 3GT expression, inhibition rate was approximately 88% (P <0.01). Empty viral vector control group, negative control group and the untreated cells siRNA group mRNA transcript levels were not significantly different (P> 0.05); immunofluorescence assay showed α1, 3GT-shRNA lentivirus transfected cells Galα (1,3 ) Gal antigen level was significantly lower than the control group (P <0.01), empty viral vector control group, negative control group and the untreated cells siRNA between groups Galα (1,3) Gal antigen expression was no significant difference (P> 0.05) . Conclusion: Through tissue explant method can achieve mouse vascular endothelial cells in primary culture and subculture, but a limited number of passages; This experiment was constructed successfully targeting α1, 3GT recombinant lentiviral vector, and using the lentiviral vector successfully α1, 3GT-shRNA expression cassette was transduced into EOMA cells to continuous expression, to achieve the targeted α1, 3GT RNA interference. Recombinant lentiviral vector effectively suppressed α1, 3GTmRNA, and inhibits its catalytic protein Galα (1,3) Gal expression. Through the experiment, the initial description of lentiviral vector-mediated RNAi technology to silence α1, 3GT gene and thus lowered xenoantigenic Galα (1,3) Gal expression strategy is feasible. So as to further study the use of RNAi technology to reduce hyperacute xenograft rejection provides an experimental basis.
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