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Transformation, Transduction and Transfection –Gene transfer methods
#5
The opening post in this thread mentions the use of nanoparticles in gene transfection. Gene therapy depends on safe and efficient gene transfer and nanoparticles are potentially useful vectors in this regard. Up until now, the vectors most recently used in gene therapy were viral-based. However, there are various issues associated with viral vectors, such as host immune responses, residual pathogenicity and limited cargo capacities of, for example, adeno-associated viruses. Nanoparticle-based approaches therefore under intense study as targeted delivery vehicles for RNA and DNA genetic drugs.

In one study in the journal Nature Materials, silver nanoparticles were developed in which a phenomenon called plasmonics was exploited. This entails resonance of nanostructured materials including gold and silver in light such that their electromagnetic fields are concentrated near the surface. This facilitates detection of the nanoprobes as it allows enhancement of the fluorescence of dyes used in their labelling. An etching technique was used to facilitate breakdown of non-internalised and potentially toxic nanoparticles for elimination. The spherical nanoparticles were also coated with a peptide that allowed them to be targeted to tumour cells, sparing healthy cells. The spherical nanoparticles were readily internalised in the tumour cells; use of these types of nanoparticles would therefore overcome the inability of RNA and DNA genetic drugs to penetrate cell membranes. The intense fluorescence of the plasmonic nanoparticles also meant that internalisation was readily detectable, while the etching technology meant that excess nanoparticles could be broken down and expelled by the kidneys.
Another potential application for gene delivery via nanoparticles would be in the area of animal genetics and breeding. One recent study described use of spherical, magnetic Fe[sub]3[/sub]O[sub]4[/sub] nanoparticles for delivery of genes expressed on DNA plasmids into the nuclei of porcine somatic cells. The nanoparticle surfaces were modified using the polycationic polyethylenimine, after which the nanoparticles showed strong binding affinity for DNA plasmids expressing the genes encoding a green (DNAGFP) or red (DNADsRed) fluorescent protein. The weight ratios of the genes to the nanoparticles could be manipulated to ensure complete binding of the DNA by the nanoparticles, even for DNA of several hundred nanometers in length. Stable and efficient co-expression of the green and red genes could be achieved in porcine kidney cells by magnetofection. The study suggested the great potential of magnetic nanoparticles for gene delivery in animal genetics and breeding studies.

References:

Friman, T., Pang, H.-B., Pallaoro, A., de Mendoza, T. H., Willmore, A.-M. A., Kotamraju, V. R., Mann, A. P., She, Z.-G., Sugahara, K. N., Reich, N. O., Teesalu, T., and Ruoslahti, E. (2014). Etchable plasmonic nanoparticle probes to image and quantify cellular internalization. Nature Materials (8 June 2014), doi:10.1038/nmat3982

Wang Y, Cui H, Li K, Sun C, Du W, Cui J, Zhao X, Chen W. A magnetic nanoparticle-based multiple-gene delivery system for transfection of porcine kidney cells. PLoS One. 2014 Jul 21;9(7):e102886. doi: 10.1371/journal.pone.0102886
 
 
 
  

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Transformation, Transduction and Transfection –Gene transfer methods00