Int J Med Sci 2007; 4(5):267-277. doi:10.7150/ijms.4.267 This issue Cite
Research Paper
1. Division of Molecular Toxicology, German Cancer Research Center, INF 280, D-69120 Heidelberg, Germany
2. Central Section for Peptide Synthesis, German Cancer Research Center, INF 580, D-69120 Heidelberg, Germany
3. Institute of Pathology, University of Heidelberg, INF 220, D-69120 Heidelberg, Germany
4. Clinical Cooperation Unit Radiation Oncology, German Cancer Research Center, INF 280, D-69120 Heidelberg, Germany
5. Department of Cell Biology, German Cancer Research Center, INF 280, D-69120 Heidelberg, Germany
6. Radiation Oncology, University of Heidelberg, INF 500, D-69120 Heidelberg, Germany
7. Division Biostatistics, German Cancer Research Center, INF 280, D-69120 Heidelberg, Germany
8. Division Biophysics of Macromolecules, German Cancer Research Center, INF 580, D-69120 Heidelberg, Germany
An efficient gene transfer into target tissues and cells is needed for safe and effective treatment of genetic diseases like cancer. In this paper, we describe the development of a transport system and show its ability for transporting plasmids. This non-viral peptide-based BioShuttle-mediated transfer system consists of a nuclear localization address sequence realizing the delivery of the plasmid phNIS-IRES-EGFP coding for two independent reporter genes into nuclei of HeLa cells. The quantification of the transfer efficiency was achieved by measurements of the sodium iodide symporter activity. EGFP gene expression was measured with Confocal Laser Scanning Microscopy and quantified with biostatistical methods by analysis of the frequency of the amplitude distribution in the CLSM images. The results demonstrate that the “BioShuttle”-Technology is an appropriate tool for an effective transfer of genetic material carried by a plasmid.
Keywords: Quantification of gene transfer, non-viral vectors, nucleus-addressed delivery, gene targeting