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최영숙,박윤정,이승진,이주연,권영민,이원규,정창평,Victor C. Yang 한국생체재료학회 2006 생체재료학회지 Vol.10 No.4
Targeted down-regulation of specific gene using antisense oligonucleotide (ODN) have gained considerable interests as therapeutic genes. The potential specificity for target gene binding and consequent inhibition of gene products makes antisense ODN attractive new class of drugs for broad clinical applications. The polyanionic charges carried by these antisense ODNs, however, present a limitation to efficient cellular uptake and consequent biological effects on gene regulation. To overcome this obstacle, a well-designed carrier system is desirable for antisense delivery. Herein the authors describe the synthesis, cellular translocation of carrier peptide and peptide-antisense ODN comprised of the cell-penetrating peptide (CPP) including TAT and protamine fragment termed as LMWP. The prepared LMWP and TAT peptide as well as their complexes with antisense ODN, for example, bcl-2 antisense ODN, were examined for cellular uptake behaviors and subcellular localization by using confocal microscopy and flow cytometry. Fluorescently tagged ODN were localized with the peptide in the cytoplasm shortly after incubation of peptide-ODN with MCF-7 breast tumor cells. The increased cell uptake achieved using the peptide compared to incubation of free ODN with cells resulted in significant down regulation of bcl-2 mRNA expression. Taken together, ODN can be delivered into the cells using cell penetrating peptides, thereby inducing marked suppression of target gene expression.