Adenovirus-mediated expression of antisense RNA transcripts complementary to pig alpha(1,3) galactosyltransferase mRNA inhibits expression of Gal alpha(1,3) Gal epitope.
OBJECTIVE
To examine the effects of the expression of antisense RNA transcripts complementary to the pig alpha(1,3) galactosyltransferase [alpha(1,3)GT]mRNA on the expression of Gal alpha(1,3) Gal structure (gal epitope) in cultured cell lines.


METHODS
Human adenoviral vectors were used to mediate the expression of antisense RNA. The expression levels of H blood group antigens and gal epitopes were analyzed by flow cytometry using FITC-UEA-I and FITC-GS-IB4 lectins, respectively.


RESULTS
Recombinant adenoviruses, Ad5anti-sGT600 and Ad5-anti-sGT1100, which express antisense RNA complementary to different regions of the pig alpha(1,3) GT mRNA, were constructed and used to infect cell line of NIH3T3. The results showed about 30% reduction in the expression level of gal epitopes on the surface of NIH3T3 cells. In addition, co-expression of human secretor type alpha(1,2) fucosyltransferase [alpha(1,2)FT]cDNA and antisense RNA complementary to the pig alpha(1,3) GT mRNA led to a further reduction in the gal epitope level.


CONCLUSIONS
Recombinant adenoviruses, Ad5anti-sGT600 and Ad5anti-sGT1100, are effective to down-regulate the gal epitope expression.
Xing
L
L
Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China.
Xia
G H
GH
Bai
X F
XF
Fei
J
J
Guo
L H
LH
D016475:3T3 Cells / Q000378:metabolism; D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D006528:Carcinoma, Hepatocellular; D002460:Cell Line; D004187:Disaccharides / Q000096:biosynthesis*; D015536:Down-Regulation; D004622:Embryo, Mammalian; D005647:Fucosyltransferases / Q000096:biosynthesis / Q000235:genetics; D005700:Galactosyltransferases / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007668:Kidney / Q000166:cytology*; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D051379:Mice; D016372:RNA, Antisense / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D004187:Disaccharides; D005647:Fucosyltransferases; D005700:Galactosyltransferases; D016372:RNA, Antisense; D012333:RNA, Messenger; D000097631:Galactoside 2-alpha-L-fucosyltransferase; C055075:galactosyl-(1-3)galactose