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Gene Therapy: Non-Viral Methods

Non-Viral Methods
Direct DNA injection is the simplest method of non-viral transfection.  (Wolff JA et al, 1990) Clinical trials have been performed successfully to inject naked DNA plasmids. Trials with naked PCR products have also been done and it had greater success. Many non-viral methods of gene transfer have been yielded by the research efforts. These method includes sonoporation, electroporation, magnetofection, receptor mediated gene transfer and gene gums are being explored.. (Wagner E et al, 1994) Ormasil is also used as a non-viral method. All methods have their own disadvantages and advantages.


Injection of Naked DNA
For the non-viral transfection it is the simplest method. Clinical trials have been done to carried out intramuscular injection of a naked DNA plasmid that have occurred with some success; though, in comparison to other transfection methods this expression has been very low. (PATIL P.M et al, 2012)


Physical Methods to Enhance Delivery

Electroporation
This is the method in which the short pulses of high voltage are used to carry DNA across the cell membrane. In the cell membrane, this shock causes a temporary pore formation allowing the molecules of DNA to pass through it. Across a wide range of cell types, electroporation is efficient in working. Though, the cell death following by electroporation limitizes its uses including clinical applications. 

Gene Gun
Another physical method of DNA transfection is the use of gene gun or particle bombardment.  DNA is coated with the particles of gold in this technique and this is loaded into a device which produces a force to achieve penetration of gold/DNA into cells e.g.,  if in the genome DNA is incorporated into wrong place, for instance in a tumor suppressor gene, it could encourage a tumour. In clinical trial it has occurred in X-SCID (X linked severe combined immunodeficiency) patients, in which by using a retrovirus hematopoietic stem cells were transduced with a corrective transgene. This leads towards the developments of T cell leukaemia in three out of 20 patients. (Wrobel I and Collins D., 1995)

Sonoporation
To deliver DNA into cells sonoporation uses ultrasonic frequencies. Acoustic caviation process is thought to disrupt cell membrane and allow the DNA to move into cells.

Magnetofection
In this method of magnetofection, DNA is complexes to the magnetic particles and underneath the tissue culture dish, a magnet is placed to bring DNA complexes into interaction with a cell monolayer.

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