Inhibition of influenza virus RNA polymerase and nucleoprotein genes expression by unmodified, phosphorothioated, and liposomally encapsulated oligonucleotides.
We have demonstrated that antisense phosphodiester (ODNs) and phosphorothioate oligonucleotides (S-ODNs) inhibit CAT (chloramphenicol acetyltransferase) protein expression in the clone 76 cell line, which is a derivative of the murine C127 cell line. This cell line expresses the influenza virus RNA polymerase and nucleoprotein (NP) genes in response to treatment with dexamethasone. Phosphodiester, phosphorothioate, and liposomally encapsulated oligonucleotides with four target sites (PB1, PB2, PA, and NP) were synthesized and tested for inhibitory effects by a CAT-ELISA assay using the clone 76 cell line. The ODNs and S-ODNs complementary to the sites of the PB2-AUG and PA-AUG initiation codons showed highly inhibitory effects. On the other hand, the inhibitory effect of the S-ODNs targeted to PB1 was considerably decreased in comparison with the other three target sites. Liposome encapsulation afforded oligomer protection in serum-containing medium and substantially improved cellular accumulation. The liposomal encapsulated oligonucleotides exhibited higher inhibitory activity than the free oligonucleotides. The activities of the unmodified oligonucleotides are effectively enhanced by using the liposomal carrier.
Hatta
T
T
Department of Industrial Chemistry, Chiba Institute of Technology, Tsudanuma, Narashino, Japan.
Nakagawa
Y
Y
Takai
K
K
Nakada
S
S
Yokota
T
T
Takaku
H
H
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D012321:DNA-Directed RNA Polymerases / Q000096:biosynthesis* / Q000235:genetics; D004337:Drug Carriers; D004797:Enzyme-Linked Immunosorbent Assay; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D005814:Genes, Viral; D008081:Liposomes; D051379:Mice; D008969:Molecular Sequence Data; D009698:Nucleoproteins / Q000096:biosynthesis* / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D009975:Orthomyxoviridae / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013873:Thionucleotides
D000998:Antiviral Agents; D004337:Drug Carriers; D008081:Liposomes; D009698:Nucleoproteins; D016376:Oligonucleotides, Antisense; D011993:Recombinant Fusion Proteins; D013873:Thionucleotides; D015500:Chloramphenicol O-Acetyltransferase; D012321:DNA-Directed RNA Polymerases