Cellular delivery of a double-stranded oligonucleotide NFκB decoy by hybridization to complementary PNA linked to a cell-penetrating peptide
2004 (English)In: Gene Therapy, ISSN 0969-7128, E-ISSN 1476-5462, Vol. 11, 1264-1272 p.Article in journal (Refereed) Published
The activation of nuclear factor B (NFB) is a key event in immune and inflammatory responses. In this study, a cell-penetrating transport peptide, transportan (TP) or its shorter analogue TP 10, was used to facilitate the cellular uptake of an NFB decoy. Peptide nucleic acid (PNA) hexamer or nonamer was linked to the transport peptide by a disulfide bond. NFB decoy oligonucleotide consisted of a double-stranded consensus sequence corresponding to the B site localized in the IL-6 gene promoter, 5'-GGGACTTTCCC-3', with a single-stranded protruding 3'-terminal sequence complementary to the PNA sequence was hybridized to the transport peptide–PNA construct. The ability of the transport peptide–PNA–NFB decoy complex to block the effect of interleukin (IL)-1-induced NFB activation and IL-6 gene expression was analyzed by electrophoretic mobility shift assay and reverse transcriptase-polymerase chain reaction in rat Rinm5F insulinoma cells. Preincubation with transport peptide–PNA–NFB decoy (1 M, 1 h) blocked IL-1-induced NFB-binding activity and significantly reduced the IL-6 mRNA expression. The same concentration of NFB decoy in the absence of transport peptide–PNA had no effect even after longer incubations. Our results showed that binding of the oligonucleotide NFB decoy to the nonamer PNA sequence resulted in a stable complex that was efficiently translocated across the plasma membrane.
Place, publisher, year, edition, pages
2004. Vol. 11, 1264-1272 p.
nuclear factor decoy, NFkappaB, PNA, transport peptide, interleukin-1beta, interleukin-6
Biological Sciences Chemical Sciences
IdentifiersURN: urn:nbn:se:su:diva-25919DOI: 10.1038/sj.gt.3302291OAI: oai:DiVA.org:su-25919DiVA: diva2:200800