Gene Therapy

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(Stem Cells)
 Hello! I’m...
GV Anika Diampoc

From BS in Accountancy. A second year student

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at Mallig Plains Colleges. Today, I presents my

report entitled “Gene Therapy (Stem Cells).
 Keypoints
01 Definition of Gene Therapy

02 Types of Gene Therapy

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03 Approaches of Gene Therapy

04 Use of Gene Therapy

Methods of Gene Therapy

05
 Definition of Gene Therapy
● Gene therapy is the introduction of genes into existing cells to prevent or
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cure a wide range of diseases.

● It is a technique for correcting defective genes responsible for disease
development.
● Gene Therapy is an experimental technique that uses genes to treat or
prevent disease.
● Gene Therapy is an experimental treatment that involves introducing
genetic material (DNA or RNA) into a person’s cells to fight disease.
 Did you know?
● On September 14, 1990 at the U.S. National Institutes of Health, W. French Anderson M.D.
and his colleagues, performed the first approved gene therapy procedure on four-year old
Ashanti DeSilva. Born with a rare genetic disease called severe combined
immunodeficiency (SCID).
● In Ashanti’s gene therapy procedure, doctors removed white blood cells from the child’s
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body let the cells grow in the laboratory, inserted the missing gene into the cells, and then
infused the genetically modified blood cells back into the patients bloodstream.
 Did you know?

Viral vectors are a tool commonly used by
molecular biologists to deliver genetic
material into cells. This process can be
performed inside a living organism (in vivo)
or in cell culture (in vitro). Viruses have
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Viruses are used as vectors to introduce the
genetic material inside the bodies. This
viruses are inactivated, they are not able to
reproduce:
● Adenoviruses

evolved specialized molecular mechanisms ● Herpes viruses DNA tumor viruses

to efficiently transport their genomes inside ● Retroviruses RNA tumor viruses
the cells they infect.
 Types of Gene Therapy
Somatic Cell Gene Therapy Germ Line Gene Therapy
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Therapeutic genes transferred into somatic Therapeutic genes transferred into the
cells. germ cells.
Eg. Introduction of genes into bone Eg. Genes introduced into eggs and
marrow cells, blood cells, skin cells, etc. sperms.
Will not be inherited later generations. It is heritable and passed on later
generations.
At present all researches directed to For safety, ethical and technical reasons, it
correct genetic defects into somatic cells. is not being attempted at present.
 Approaches of Gene Therapy
1. In Vivo Gene Therapy: direct delivery of genes into the cells of a
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particular tissue in the body.

2. Ex Vivo Gene Therapy: transfer of genes to cultured cells and

reinsertion.
 In Vivo Gene
● Therapy
Direct delivery of therapeutic gene into target
cell into patients body.
● Carried out by viral or non viral vector systems.
● It can be the only possible option in patients
where individual cells cannot be cultured in
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vitro in sufficient numbers (e.g. brain cells)
● In vivo gene transfer is necessary when
cultured cells cannot be re-implanted in
patients effectively.
● Example: In patients with cystic fibrosis, a
protein called cystic fibrosis transmembrane
regulator (CFTR) is absent due to a gene defect
Ex Vivo Gene
Therapy
● Isolate cells with genetic defect from a
patient.
● Grow the cells in culture.
● Introduce the therapeutic genes.
● Select genetically corrected cells and
grow.
● Transplant the modified cells to the
patient.
● Example: 1st gene therapy – to correct
deficiency off enzyme, Adenosine
deaminase (ADA); Caused due to defect
in gene coding for ADA.
 How it Works?
A vector delivers the therapeutic gene into
a patients target cell. The target cells
become infected with the viral vector. The
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vector’s genetic material is inserted into

the target cell. Functional proteins are
created from the therapeutic gene causing
the cell to return to a normal state.
 Vectors in Gene Therapy

Vectors Two Main

● To transfer the desired ● ViralClasses
Vectors
gene into a target cell, a ● Non viral Vectors
carrier is required. Such
vehicles of gene delivery
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are known as vectors

 Viral Vectors
Retrovirus Vector
● System
The recombinant retroviruses have the
ability to integrate into host genome in a
stable fashion.
● Can carry a DNA of size – less than 3.4 kb
● Replication defective virus particles
● Target cell - dividing
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Adeno Associated Virus
● It is a human Vector
virus that can integrate
into chromosome 19.
● It is a single stranded, non pathogenic
small DNA virus.
● AAV enters host cell, becomes double
stranded and gets integrated into
chromosome.
Adeno Virus Vector
● Adeno virusSystem
with a DNA genome
– good vectors.
● Target – non dividing human cell.
● E.g. Common cold adenovirus
Herpex Simplex Virus Vector
● Viruses which have natural
tendency to infect a particular
type of cell.
● They infect and persist in nervous
cells.
 Non Viral Vector
Pure DNA Construct Lipoplexes
● Direct introduction of pure DNA construct ● Lipid DNA complexes;
into target tissue. DNAconstruct surrounded by
● Efficiency of DNA uptake by cells and artificial lipid layer.
expression rather low. ● Most of it gets degraded by
● Consequently, large quantities of DNA lysosomes.
have to be injected periodically.
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DNA Molecular Human Artificial

Can carry a large DNA i.e.,
CommonlyConjugates Chromosome
●
● used synthetic conjugate is
with one or more therapeutic
poly – L – lysine bound to specific target
genes with regulatory
cell receptor.
elements
● Therapeutic DNA is then made to combine
with the conjugate to form a complex.
● It avoids lysosomal breakdown of DNA.
 What Gene Therapy can Achieve
1. Replacing a mutated gene that causes disease with a healthy copy
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of the gene
2. Inactivating, or “knocking off,” a mutated gene that is functioning
improperly.
3. Introducing a new gene into the body to help fight a disease.
 Uses of Gene Therapy
1. Replace missing or defective genes;
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2. Deliver genes that speed the destruction of cancer cells;

3. Supply genes that cause cancer cells to revert back to normal cells;
4. Deliver bacterial or viral genes as a form of vaccination;
5. Provide genes that promote or impede the growth of new tissue;
6. Deliver genes that stimulate the healing of damaged tissue.
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 Methods of Gene Delivery
 Microinjection
 Gene Gun
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 Process of using a glass micropipette to

 Employs a high-pressure
delivery system to shoot
tissue with gold or tungsten
particles that are coated
with DNA.
insert microscopic substances into a
single living cell.
 Normally performed under a specialized
optical microscope setup called a
micromanipulator.
 Other Types of Gene Therapy
Gene Augmentation Therapy
● Most common form of gene therapy

● Foreign gene replaces missing or defective gene.

● E.g., Replacement of defective p53 gene by a normal one in liver cancer.

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Gene Inhibition Therapy

● Done to block the overproduction of some proteins.

● Two Types – Antigene and antisense therapy

 Antigene – blocks transcription using antigene oligonucleotide.

 Antisense – blocks translation using antisense oligonucleotide.

 Goal of Gene Therapy
● A normal gene may be ● An abnormal gene could
inserted into a non-specific be swapped for a normal
location within the genome gene through homologous
to replace a non-functional recombination
gene. This approach is the
most common
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● The abnormal gene could ● The regulation (degree to

be repaired through which a gene is turned on
selective reverse mutation, or off) of a particular
which returns the gene to gene could be altered
its normal function.
 Very interesting facts!
Genes are Medicine.
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Gene Therapy is “the use of genes as medicine.” It involves the

transfer of a therapeutic or working gene copy into specific cells of an
individual in order to repair a faulty gene copy. Thus, it maybe used to
replace a faulty gene, or to introduce a new gen whose function is to
cure or to favorably modify the clinical course of a condition.
Thank you!
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