tailieunhanh - Báo cáo khoa học: Cellular uptake of antisense oligonucleotides after complexing or conjugation with cell-penetrating model peptides

The uptake by mammalian cells of phosphorothioate oli-gonucleotides was compared with that of their respective complexes or conjugates with cationic, cell-penetrating model peptides of varying helix-forming propensity and amphipathicity. An HPLC-based protocol for the synthesis and purification of disulfide bridged conjugates in the 10– 100 nmol range was developed. Confocal laser scanning microscopy (CLSM) in combination with gel-capillary electrophoresis and laser induced fluorescence detection (GCE-LIF) revealed cytoplasmic and nuclear accumula-tion in all cases. . | Eur. J. Biochem. 269 4025-4032 2002 FEBS 2002 doi Cellular uptake of antisense oligonucleotides after complexing or conjugation with cell-penetrating model peptides J. Oehlke P. Birth E. Klauschenz B. Wiesner M. Beyermann A. Oksche and M. Bienert Institute of Molecular Pharmacology Berlin Germany The uptake by mammalian cells of phosphorothioate oligonucleotides was compared with that of their respective complexes or conjugates with cationic cell-penetrating model peptides of varying helix-forming propensity and amphipathicity. An HPLC-based protocol for the synthesis and purification of disulfide bridged conjugates in the 10100 nmol range was developed. Confocal laser scanning microscopy CLSM in combination with gel-capillary electrophoresis and laser induced fluorescence detection GCE-LIF revealed cytoplasmic and nuclear accumulation in all cases. The uptake differences between naked oligonucleotides and their respective peptide complexes or conjugates were generally confined to one order of magnitude. No significant influence of the structural properties of the peptide components upon cellular uptake was found. Our results question the common belief that the increased biological activity of oligonucleotides after derivatization with membrane permeable peptides may be primarily due to improved membrane translocation. Keywords oligonucleotide-peptide conjugates cellular uptake cell-penetrating peptides. The effectiveness of antisense oligonucleotides and peptide nucleic acids in modifying mammalian cell function can be improved substantially by covalent attachment or complexing with natural cell-penetrating peptide sequences 1-4 . This increase in biological activity has been commonly attributed to an enhanced cellular uptake of the conjugates 5-7 . The peptide components used to date have been protein-derived sequences that exhibit very different structural properties ranging from lipophilic to unstructured and highly positively .

• Báo cáo khoa học
• Report medicine
• traditional medicine
• scientific reports
• intermediate cells
• cell proliferation
• medical analysis
• thesis report
• medical report
• medical knowledge
• medical research
• analysis of cytoplasmic
• environmental medicine
• raccoon experiments
• Content and Language Integrated Learning
• Analysis of term extraction for English
• English medical texts
• Vietnamese medical experts
• Educational philosophy across Europe
• Medical image analysis
• Computer aided diagnosis
• FLAIR magnetic resonance imaging
• Realistic lesion insertion
• Medical data augmentation
• Diffuse lung diseases
• Tomosynthesis imaging
• Spinal abnormalities
• Health informatics
• Medical microwave imaging
• statements and documents scientific reports
• medical reports
• clinical trials
• medical management
• health care
• scientific research
• health policy
• pathology
• anatomy
• health education