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Gene Transfer Technologies: T.Manoj Kumar
Gene Transfer Technologies: T.Manoj Kumar
T.Manoj Kumar
What is gene therapy ????
• Gene therapy is an experimental technique that uses genes to
treat or prevent disease.
• In 1990’s, first approved gene therapy case in The United States took
place on 14th September 1990, at the national institute of health, under
the direction of professor William French Anderson.
• Using a vector virus the PDX-1 gene has been transferred (into
mice) where the gene is expressed by pancreatic cells which now
produce insulin.
divided as,
• Non-viral methods.
• Viral methods.
Non-Viral Delivery Systems
Limitations:
1. Method is fast.
2. Less costly.
1. Costly.
• Methods may prove useful for gene transfer into cereals which
do not regenerate from cultured cell easily.
• Technically simple.
Limitations
1. Less specific.
2. Less efficient.
• The coated beads are then attached to the end of the plastic
bullet and loaded into the firing chamber of the gene gun.
• An explosive force fires the bullet down the barrel of the gun
towards the target cells that lie just beyond the end of the
barrel.
• When the bullet reaches the end of the barrel it is caught and
stopped, but the DNA coated beads continue on toward the
target cells.
• Some of the beads pass through the cell wall into the
cytoplasm of the target cells.
• Here the bead and the DNA dissociate and the cells become
transformed.
• Once inside the target cells, the DNA is solubilised and may
be expressed.
original 22-caliber biolistic gun
• The particle gun has also been used with pollen, early stage
embryoids, meristems and somatic embryos.
Advantages and limitations of biolistics
Limitations
1. Integration is random.
2. Requirement of equipments.
LIPOSOME MEDIATED GENE TRANSFER
• These are artificial vesicles that can act as delivery agents for
exogenous materials including transgenes.
1. Simplicity.
2. Long term stability.
3. Low toxicity.
4. Protection of nucleic acid from degradation.
CALCIUM PHOSPHATE MEDIATED DNA TRANSFER
• DNA can be transferred with the help of DAE Dextran also. DAE-
Dextran may be used in the transfection medium in which DNA is
present.
1. Retrovirus (RV).
2. Adenovirus (AV).
3. Adeno Associated Virus (AAV).
Retro Virus Vectors
Properties:
Features
• Lack envelope.
• http://encarta.msn.com/media_461561269/Gene_Therapy.html