[The porcine alpha1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte negative express GT].

摘要: OBJECTIVE: To study whether the porcine alpha1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte negatively expresses GT mRNA and resists to cytotoxicity of nature antibody in human serum. METHODS: The vector (pPNTloxPGTsiRNA) were construct with pPNTloxPGT pMXSV/U6 vector. Positive-negative selection was used produce a pPNTloxPGTsiRNA knockout (+/-) clone. expressions detected northern blot. Complement-mediated NAb after incubation hepatocytes NAbs complement determined using 3- (4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS, tetrazolium salt) colorimetric assay. RESULTS: negative express mRNA. Only 14% 18% can be at highest serum concentration. just as wild type cells are 77% 83%. CONCLUSION: a1, resisted