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
in Chinese
| WPRIM
| ID: wpr-250189
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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Swine
/
Killer Cells, Natural
/
Transfection
/
Cells, Cultured
/
Cloning, Molecular
/
Gene Targeting
/
Gene Silencing
/
Hepatocytes
/
Cell Biology
/
Cytotoxicity, Immunologic
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Hepatology
Year:
2004
Type:
Article
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