Targeting of antisense PNA oligomers to human galanin receptor type 1 mRNA
作者： Kalle Kilk , Anna Elmquist , Külliki Saar , Margus Pooga , Tiit Land
DOI: 10.1016/J.NPEP.2004.06.005
关键词: RNA 、 Molecular biology 、 Biochemistry 、 Cell-penetrating peptide 、 Galanin 、 Oligonucleotide 、 Nucleic acid 、 Peptide 、 Biology 、 Galanin receptor 、 Peptide nucleic acid
摘要: In this work, we have targeted positions 18-38 of the human galanin receptor type 1 (GalR1) mRNA coding sequence with different peptide nucleic acid (PNA) oligomers. This region has previously been shown to be a good antisense and therefore aimed identify subregions and/or thermodynamic parameters determining efficacy. Nine PNA oligomers were conjugated cell-penetrating peptide, transportan, enhance their cellular uptake. Concentration-dependent down-regulation GalR1 protein expression in melanoma cell line Bowes was measured by radioligand binding assay. No reduction level observed upon treatment, thus, effect concluded translational arrest. Judging from EC50 values, targeting regions 24-38 (EC50=70 nM) or 27-38 (EC50=80 most potent suppressors expression. parameter predicted M-fold algorithm found correlate activities. Presence some not increase efficiency PNA. short unpaired triplet between nucleotides 33 35 target was, on other hand, critical for efficient down-regulation. Thus, results are high impact designing Specific study demonstrate 20-fold more as achieved before.
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