Gene Therapy Types Applications Regards- urvashi shakarwal

1. Urvashi Shakarwal
M Pharm Pharmacology 1
Presented by-
Urvashi Shakarwal
M Pharm Pharmacology
Amity Institute of pharmacy
Amity University
Lucknow U.P.

2. Urvashi Shakarwal
M Pharm Pharmacology
Introduction
Gene therapy is to transfer a gene from one DNA molecule to another DNA
molecule. It is the insertion of genes into the individual’s cell and tissues to
treat a disease.
The directed desirable gene transfer from one organism to another and the
subsequent stable integration and expression of foreign gene into the genome
is referred as genetic transformation.
Unstable transformation occur when DNA is not integrated into host genome.
Stable transformation occur when DNA is integrated into host genome and is
inherited in subsequent generations.
The transferred gene is known as transgene and the organism that develop
after a successful gene transfer is known as transgenic organism.
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3. Urvashi Shakarwal
M Pharm Pharmacology
Steps involved in gene therapy
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4. Urvashi Shakarwal
M Pharm Pharmacology
Approaches involved in gene therapy
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M Pharm Pharmacology 5

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M Pharm Pharmacology 6

7. Urvashi Shakarwal
M Pharm Pharmacology
Vectors involved in gene therapy
To transfer the desired gene into a target cell a carrier is required.
Such vehicle of gene delivery are known as vectors.
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8. Urvashi Shakarwal
M Pharm Pharmacology
Viral vectors
Viral vectors are mostly derivatives of viruses that infects animals.
These viral vector can be divided into 2 groups:
1.Free replicating viruses- that multiply within the cell, but do not
integrate into the genome of the host.
2.Integrateing viruses-that can integrates into the host genome.
They enters the cell, copy their RNA genome into DNA.
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9. Urvashi Shakarwal
M Pharm Pharmacology
Types of viral vectors
Retrovirus vector system
They uses the enzyme reverse transcriptase to convert
the RNA into DNA which is integrated into gemone
further enters the host cell.
It have the ability to infect a wide variety of cell type
with high efficiency.
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10. Urvashi Shakarwal
M Pharm Pharmacology
Adenovirus vector system
The AAV genome comprises of inverted terminal repeats (ITR)
as both ends of the DNA stand and open reading frames.(ORFS).
This type of viruses is being used, because it is non pathogenic
and do not build an immune response.
AVV vectors are used to deliver genes to the brain, this is
possible because AAV viruses can infect non-dividing cells such
as neurons.
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11. Urvashi Shakarwal
M Pharm Pharmacology
Herpes simplex viruses
Herpes viruses includes herpes simplex viruses
that rarely can cause encephalitis and infect the
non dividing cells so it have the ability to
transduce neurons. Its advantage is being able to
infect non dividing cells that help in treating
neurological disorders.
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12. Urvashi Shakarwal
M Pharm Pharmacology
Advantages of viral vectors
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13. Urvashi Shakarwal
M Pharm Pharmacology
Non viral vector system
Physical methods-
1.Electroporation
2.Microinjection
3.Gene gun or a biolistic particle delivery system
4. Magnetofection
Chemical methods-
1.DNA transfer by calcium phosphate method
2.Liposome medicated transfer
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14. Urvashi Shakarwal
M Pharm Pharmacology
Electroporation
Electroporation is a microbiology technique in which an electrical field is applied to
cells in order to increase the permeability of the cell membrane allowing chemicals,
drugs, or DNA to be introduced into the cell.
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15. Urvashi Shakarwal
M Pharm Pharmacology
Microinjection
The microinjection is the process of transferring the desirable
DNA into the living cell, through the use of glass micropipette.
Glass micropipette is usually of 0.5 to 5 micrometer.
It easily get penetrates into the cell membrane and nuclear
envelope. The desired gene is then injected into the sub cellular
compartment and needle is removed.
Limitations of microinjection
-Costly
-Skilled person required
-More useful for animal cells.
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16. Urvashi Shakarwal
M Pharm Pharmacology
Gene gun or a biolistic particle delivery
system
1.Biollistics or particle bombardment also known as gene gun
technique is a physical method that uses accelerated micro projectiles
to deliver DNA or other molecule into intact tissues and cells.
2.The gene gun is a device that literally fit DNA into target cells.
3.The DNA to be transformed into the cell which are coated onto
microscopic beads made of either gold or tungsten.
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17. Urvashi Shakarwal
M Pharm Pharmacology
4.The coated beads are then attached to the end of the plastic bullets and
loaded into the firing chamber of the gene gun.
5.An explosive force fires the bullet with DNA coated beads towards the
target cells that lie just beyond the end of the barrel.
6.Some of the beads pass through the cell wall into the cytoplasm of the
target cells.
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18. Urvashi Shakarwal
M Pharm Pharmacology
Magnetofection
Magnetofection is a simple and highly efficient transfection
method that uses magnetic fields to concentrate particles
containing nucleic acid into the target cells.
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19. Urvashi Shakarwal
M Pharm Pharmacology
Chemical methods
1.DNA transfer by calcium phosphate method
The process of transfection involves the mixture of isolated DNA with
solution of calcium chloride and potassium phosphate.
Cell are then incubated with precipitated DNA either in solution or in
tissue culture dish.
A fraction of cells will take up the calcium phosphate DNA precipitate
by endocytosis.
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20. Urvashi Shakarwal
M Pharm Pharmacology
2.Liposome medicated transfer
Liposome are spheres of lipids which can be used to transport
molecules into cells.
These are artificial vesicles that can act as delivery agents for
exogenous materials including transgenes.
Promote transport after fusing with the cell membrane.
Cationic lipids are those having a positive charge are used for the
transfer of nucleic acid.
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21. Urvashi Shakarwal
M Pharm Pharmacology
Various strategies for gene therapy
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M Pharm Pharmacology
Applications of Gene Therapy
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M Pharm Pharmacology 23

24. Urvashi Shakarwal
M Pharm Pharmacology
Chronic Granulomatus Disorder (CGD)
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M Pharm Pharmacology
Hemophilia
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M Pharm Pharmacology
Gene therapy trial for inherited
blindness
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M Pharm Pharmacology
Gene therapy for acquired diseases
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M Pharm Pharmacology
Other acquired diseases
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29. Urvashi Shakarwal
M Pharm Pharmacology
Uses of gene therapy
Clinical gene transfer applications
Vaccine development
Production of transgenic animals
Treatment of cancer and AIDS
Gene discovery
Gene therapy
Enhancing the resistance of plant
Genetically modified organism
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M Pharm Pharmacology 30