BIOL 103 |
| In the past five years, gene therapy has made significant progress. Gene therapy provides the potential to cure various diseases by replacing a missing gene or introducing a functional substitute of a defective gene. The main challenge of gene therapy has always been to find an effective means of gene transfer. We are exploring the delivery of plasmid DNA based on the cell-surface receptor for RGD. A peptide containing RGD sequence was synthesized and conjugated with plasmid DNA encoding the firefly luciferase gene by using diazocoupling method to achieve covalent modification of the DNA backbone. The conjugates was tested to make sure they maintain the functionality of the expression cassette. After modification, interaction between the functional domains on plasmid DNA and the extracelluar receptors may enhance cell-specific targeting and increase plasmid uptake through receptor-mediated endocytosis. In addition, the plasmid DNA must be neutralized and compacted into a small vesicle that can be easily taken by the cells. The plasmid-peptide conjugations are neutralized and compacted by such transfection reagents as Fugene 6, low molecular weight polylysine, high molecular weight polylysine and cell permeable adenine-thymine-binding fluorescent dye Hoechst 33285. The ability of the RGD peptide to enhance gene delivery has been tested in cellular models. The transfection data indicates that the ligand modification can improve the tranfection efficiency. |
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Chemistry and Metabolism
4:30 PM-6:30 PM, Tuesday, 12 September 2006 Moscone Center -- Hall D, Poster
Division of Biological Chemistry |