Gene therapy is the study of methods thrugh which gene can be transferred from ons cell to another which can aid in treating thousands of disorders

1. C
GENE THERAPY
Presented By-Prachi mehta
M pharm 1st year
Guided by- Dr Bhrigu K Das
SCHOOL OF PHARMACEUTICAL SCIENCE
GCU Guwahati

2. CONTENTS
• GENE THERAPY IN BREIF
• TYPES OF GENE TRANSFER
• ADVANCEMENT IN GENE THERAPY
• ETHICAL ISSUES
• SUMMARY

3. DEFINATION
• GENE THERAPY is replacement of defected or absent gene with the correct one.
• It is the insertion of gene into an individual’s cell and tissue to treat a disease or
hereditary disease where a mutant allele is replaced by a function of gene.
• Types of diseases gene therapy can treat – Haemophilia, Sickle cell anaemia ,
Parkinsons disease, Chronic Granulomatous disease , Cancer , Diabetes, Alzehimers
disease.
• Disease that led to the discovery of gene therapy – ADA-SCID

4. ADA-SCID[adenosine deaminase]
• GENE therapy was done for the first time in a 4 year old girl.
• Problem- gene for ADA was absent.
• It is required for maturation of T lymphocytes, this results in SCID which is severe
combined immuno deficiency. This results in two things :
• FIRST- Amount of circulating T lymphocytes decrease due to a high amount of toxic
metabolites circulating through the body affecting thymus.
• SECOND- survival rate of T cells produced decreases.
• I am not having fun bro, study hard and release me

5. Bubbly boy disease

6. Process of gene therapy for ada

7. TYPES OF GENE THERAPY
• Depending on the type of cell you are performing gene therapy
• 1. somatic gene therapy
• 2. germ line gene therapy
Gene
therapy
Somatic
gene therapy
Germ line
gene therapy

8. DIFFERENCE
Somatic gene therapy
• Most commonly
practiced
• This does not get
transferred to further
progeny
Germ line gene
therapy
• Very rarely practiced
• This can get
transferred to further
progeny, has many
ethical issues.

9. APPROACHES
• EX VIVO GENE THERAPY
• IN VIVO GENE THERAPY

10. EX VIVO GENE THERAPY

11. IN VIVO GENE THERAPY
• Involves introduction of therapeutic DNA directly into patient’s body. DNA is introduced
• by cell specific direct injection into tissue in need.
• DNA in the form of plasmid vector is introduced by
• A dermal vaccination. Once incide the body and in
• Contact with specifically targeted cells, the inserted
• DNA is incorporated in tissue cell.

12. METHODS of GENE THERAPY
• PHYSICAL METHOD
• 1. gene gun method
• 2. Micro injection
• CHEMICAL METHOD
• 1. Detergent mixture
• 2. lipofection
VECTOR METHOD
1. ADENO VIRUS 2. RETRO VIRUS 3. HERPES SIMPLEX VIRUS

13. Virus with thier features
Retro virus
• Can integrate DNA efficiently into the host genome
• DNA production replication.
• It is made of RNA
• RETRO RECEPTOR ,3-9 kb
Adeno virus
• DOUBLE stranded Dna genome
• Highly target specific and has controlled integration
• Relatively easy to culture and produce in large number
• CAR RECEPTOR, Small size allows to enter many body tissue ,5 - 40kb
Herpesssimplex
virus
• Higher DNA carrying capacuty
• Suitable for invivo gene therapy 120 – 200 kb
• difficult to produce , HSPG receptor

14. VIRUS WITH THEIR RECEPTORS

15. Gene gun method
• Also known as biolistic gene gun method. It introduces DNA into skin bound into small
gold particles for this purpose DNA is precipitated onto gold microparticles which are
then loaded into plastic loading one and half piece of tubing are cut and loaded into
the gene gun . The velocity and the size of the gold particle determines the depth of
the penetration into skin layers .
Precipitate
DNA
Load to
tubing
Rotate to
coat tube and
cut tube
Load to the
gene gun
Deliver to
targeted cells

16. • ADVANTAGE – can be used in any type of cell.
• Good for muscle and tumor treatment gets higher expression then needles,
• DISADVANTAGE- damage of a significant number of cells

17. Biolistic method

18. MICROINJECTION
• The transformation method of microinjection is particularly efficient when inserting DNA into
giant cells . The microinjection technique introduces DNA into animal cells ( eggs , oocytes ,
and embryos) or plant protoplast using a micropipete( fine tipped glass needle ). this process
involves incorporating DNA straight into cytoplasm or nucleus.
• ADVANTAGE- can be used to treat a small number of cells.
• Selection of transformed cells is easy.
• DISADVANTAGE- Time consuming method
• Requires specialised and computerised microscope setup.

20. CHEMICAL METHODS
• DETERGENT MIXTURES- calcium phosphate , dextran is mixed with functional cDNA of
desired function.
• The mixture is introduced near the vicinity of recipient cells .
• the chemical disturbs the cell membrane , widens the pore size and allows Cdna to pass
through the cell.
• LIPOFECTION – It is a technique used to inject genetic materials into a cell by means of
liposomes
• Liposomes are artificial phospholipid vesicles used to deliver a variety of molecules including
DNA into the cells .

21. Chemical detergent vs lipofection
one breaks the membrane while other works as a carrier.

22. ETHICAL ISSUES
• 1. interfering with gods plan
• 2. leads to eugenics
• 3. is it alright to use gene therapy in babies in prenatal stage .

23. Advancement
• in a new gene therapy method developed by university of Florida in jan 2012 ,
researchers found treatment for a common form of blindness ( X linked retinitis
pigmentosa ) that strikes youngsters and adult.
• A gene therapy called NLX P101dramatically reduces movement impairment in
parkinsons patient, according to results of phase 2 study .

24. Summary and questions
• We learnt the history of gene therapy , how was the first case of gene therapy solved .
• Methods of gene transfer using vectors , physical method , chemical method.
• Q1. What is gene therapy and how did it play its role in ADA ?
• Q2. explain any 2 gene therapy method.
• Q3. write any 3 viral vectors used in gene therapy with their receptor.
• Q4. Why do we use gold or tungsten in gene gun method ?

25. Thank you .
• FINALLY DONE , HAPPY LEARNING
• NAP TIME .