“For an effective transfer of genetic material carried by a plasmid, “BioShuttle” technology is an appropriate tool” the results demonstrated by a study.

BioShuttle

The beginning of cancer can be described as a genetic accident. More than 200 genes play a role in the generation of cancer. The genetic malfunction can be compensated by introducing one or more therapeutic genes in to the target cells for the treatment of genetic diseases. For the efficient transport of therapeutic genetic material in to the nucleus of the target cells are the direct passage through the nuclear envelope in to the nucleus and the quick transport across the plasma membrane in to the cytosol. To overcome these barriers numerous delivery systems have been developed such as viral vectors, ultrasound and non-viral peptide based carrier systems.

The modules of “Clamp PNA Bioshuttle” consist of:

  • As mediator of the transport across outer membranes, a transmembrane transport unit is present.
  • Mediator of the cellular compartment addressed transport.
  • Hybridization-site harboring two identical PNA-sequences connected with lysine via a duplex glycine spacer.

‘Endocytosis’ is required for the mechanism of lipid-mediated transfection. The efficacy of lipofection is limited and the unprotected DNA in the cytoplasm could be degraded by resident cytosolic DNAses. But the mechanism of the BioShuttle mediated gene transfer remains unclear.  With the help of the reporter gene EGFP the efficiency between BioShuttle and lipofection was compared. While most cells showed a diffuse signal, a strong fluorescent signal was found in few HeLa-cells transfected by lipofection.  But BioShuttle method is completely different from lipofection as there is a clear fluorescence signal in nearly all the cells with a cytoplasmic signal higher than the cell nucleus.

An appropriate standard area serving as a basis for the calculation of frequency and intensity of the signals had developed because of the difference of the fluorescence intensities in lipofection treated cells.

A solution for a quantification of gene expression after transfer of phNIS-IRES-EGFP can be contributed by the use of physical and bio-mathematical knowledge. The microscopic epifluorescence method was not suitable for an examination of the transfection according to the EGFP experiments with BioShuttle-mediated gene transfer. For an accurate evaluation of the efficiency of the transcription demands new techniques like computer aided microscopy.

It is difficult for the quantitative detection of the expression of therapeutic or reporter genes. In addition to that the uptake of the different amounts of genetic material in to the target cells and the different gene expression followed include a reliable quantification. The quantification of the intracellular gene expression rate after gene transfer can be accounted by the equations of the numeric values of light intensities and their corresponding frequencies. The bio-statistical method used is able to recognize the cellular distribution of the different gene expression. The direct comparison of the efficacy of different gene transfer procedures are allowed by this method.

For monitoring the gene transfer efficiency and the controlled gene expression in the target tissue new strategies are needed for the new development of gene delivery system. A promising solution is represented by the monitoring improvements in “transfection via confocal laser scanning microscopy (CLSM)” and its mathematical evaluation. Universal gene delivery system was not identified is an unquestionable fact. For the genetic materials of the cell Bio-shuttle can be considered as a smuggler. The BioShuttle targeting system might offer a successful step in the cell type-exact delivery of molecules for the intervention by RNAi, aptamers and ribozymes. But the gene therapy has to go a long way.

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