The porcine alpha1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte negative express GT / 中华肝脏病杂志
Chinese Journal of Hepatology
; (12): 482-484, 2004.
Article
Dans Zh
| WPRIM
| ID: wpr-250189
Responsable en Bibliothèque :
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To study whether the porcine alpha1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte negatively expresses GT mRNA and resists to the cytotoxicity of nature antibody in human serum.</p><p><b>METHODS</b>The porcine alpha1, 3 galactosyltransferase gene siRNA targeted vector (pPNTloxPGTsiRNA) were construct with pPNTloxPGT and pMXSV/U6 vector. Positive-negative selection was used to produce a heterozygous pPNTloxPGTsiRNA knockout (+/-) clone. The GT mRNA expressions were detected with northern blot. Complement-mediated NAb cytotoxicity after incubation of hepatocytes with NAbs and complement was determined using 3- (4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS, tetrazolium salt) colorimetric assay.</p><p><b>RESULTS</b>The pPNTloxPGTsiRNA targeted porcine hepatocyte (+/-) negative express GT mRNA. Only 14% to 18% cytotoxicity can be detected at the highest serum concentration. The pPNTloxPGT targeted porcine hepatocyte (+/-) express GT mRNA just as the wild type porcine cells and the cytotoxicity are 77% to 83%.</p><p><b>CONCLUSION</b>The porcine a1, 3 galactosyltransferase gene siRNA targeted heterozygous hepatocyte (+/-) negative express GT and resisted to nature antibody in human serum.</p>
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Suidae
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Cellules tueuses naturelles
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Transfection
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Cellules cultivées
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Clonage moléculaire
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Ciblage de gène
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Extinction de l'expression des gènes
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Hépatocytes
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Biologie cellulaire
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Cytotoxicité immunologique
Limites du sujet:
Animals
langue:
Zh
Texte intégral:
Chinese Journal of Hepatology
Année:
2004
Type:
Article