Methods Of Transfection

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Current research methods of transfection, delivering foreign DNA into cells, have capitalized on using non-viral vectors because of the recent advantages researchers have been able to exploit. The process of transfecting cells runs into a number of problems by way of the cell’s own defense mechanisms. Vectors must be able to not only enter the cell past the cell’s membrane but also must be able to make its way into the cell’s nucleus to access the targeted genetic material. The problem with traditional transfection methods is that they are not able to enter the cell in high efficiency without triggering an immune response. This, coupled with the inability for prolonged gene expression in vivo even once transfected, results in a very expensive and ineffective method for introducing a foreign plasmid into the cell. In the past viral vectors had been used with a degree of success in vitro, but because they lack a high degree transfection efficiency and duration of gene expression using them for transfection could not produce substantial practical applications. Another problem is that these laboratory-engineered viruses had low success rates in vivo due to activating an immune response. New techniques are being discovered by modifying non-viral vectors in novel ways, producing increasingly effective methods for transporting DNA into cells with the hope of clinical application and advancing gene therapy.
Research done by Figueroa et al. (2013) has turned up an advance in the method of non-viral gene delivery utilizing gold nanoparticles (AuNP). In their work they have developed an exceptionally efficient way of transfection by experimentally improving certain desirable traits of the gold nanoparticle polyamidoamine (AuPAMAM) c...

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... of effectiveness when mTat and PEI are combined to take advantage of each vector’s positive traits for transfection. Cytotoxicity results from the mice transfected in vivo found that there was no significant difference in cell viability between mTat, PEI, or mTat/PEI complex. These results demonstrated the remarkable ability to significantly increase both the efficiency of transfection and gene expression in vivo without conferring any increase in cytotoxicity. Cell penetrating peptides (CPP), including mTat, have proven to be effective at crossing cell membranes quickly and are able to enter the cell based on their small and amphipathic nature (Bolhassani, 2011). By controlling the cargo CPP carries and the target it delivers it to, these carriers may hold a myriad of uses in vivo including selectively disrupting cancer cells and other gene therapy techniques.

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