VIDYALAYA,

TIKIRA, GHATAGAON, KEONJHAR - 758029

SESSION 2022-23
BIOLOGY INVESTIGATORY
PROJECT REPORT
TOPIC :- GENE THERAPY
SUBMITTED BY :-
Mr. LOKESH RANJAN MAHANTA
CLASS : XII-SCIENCE SUBMITTED TO :-
SECTION : ‘B’
SCHOOL ROLL NO. : 09 Mr. AMAN RATHOD
CBSE ROLL NO. : (PGT Biology)
SUBJECT : BIOLOGY
 CERTIFICATE
This is to certify that this Biology Investigatory Project on the topic
‘GENE THERAPY’
has been successfully completed by
MR. LOKESH RANJAN MAHANTA
of class XII(B) under the guidance of
MR. AMAN RATHOD
for the fulfillment of the curriculum of the Central Board of Secondary
Education (CBSE) leading to 12th Board examination of session 2022-23.

i
Teacher In-charge External Examiner Principal

Date : / /
 ACKNOWLEDGMENT
In the accomplishment of this project successfully, I would like to
express my gratitude and appreciation to all those who gave me the
possibility to complete this.
Primarily I would thank God for being able to complete this
project with success. Then I would like to thank our Biology teacher
Mr. Aman Rathod whose valuable guidance and pieces of advice
have been the ones that helped me patch this project and make it a
full-proof success. I would also like to thank our Principal Mr.
Suvendu Kumar Moharaj for his suggestions and his instructions
that served as the major contributor towards the completion of the
project.
Finally, I would like to thank my parents for helping me
economically and my classmates who have helped me with their ii

valuable suggestions that have been helpful in various phases of the

completion of the project.
 DECLARATION
I hereby declare that the investigatory project report
entitled ‘GENE THERAPY’ submitted to Odisha Adarsha
Vidyalaya, Tikira, is a record of an original work done by me
under the guidance of Mr. Aman Rathod, PGT Biology,
Odisha Adarsha Vidyalaya, Tikira, and this project report is
submitted for the partial fulfillment in particular fulfillment of
curriculum of Central Board of Secondary Education (CBSE)
leading to the 12th Board examination of session 2022-23. This
project report is not submitted to any other educational institute
or for any other purpose.
iii
CLASS : XII(B)
 CONTENTS
1. Certificate ……………..……………………………………….. i
2.Acknowledgement ………………………………………….. ii
3.Declaration ……………………………………………………. iii
4.Introduction …………………………………………………..... 2
5. Brief history of Gene Therapy …………………………… 4
6. Types of Gene Therapy ……………………………….......... 5
7. Vectors of Gene Therapy …………………………………. 13
8.Methods of Gene Delivery ………………………………… 20
9. Success cases of Gene Therapy …………………………. 22
10.Problems with Gene Therapy ………………………….. 23
11.Ethical issues ……………………………………………......... 24
12.Conclusion ………………………………………………….… 25
13.Bibliography ……………………………………………......... 26
 INTRODUCTION

WHAT IS A GENE ?
A gene is a basic unit of heredity.
It is located in chromosomes.
It encodes protein formation.
Replication
Transcription
Translation
DNAReverse transcriptionRNA Protein

Proteins formed to carry out

various functions.
If gene dysfunctions, it causes
altered protein formation.
When there is a mutation in the
2
gene, then the wrong codon will
code for the wrong amino acid in the
protein which leads to an alteration
in the basic structure of the protein.
WHAT IS GENE THERAPY?

Gene therapy is a medical approach

that treats or prevents diseases by
correcting the underlying genetic
problem.

These techniques allow doctors to

treat a disorder by altering a
person’s genetic makeup instead of
using drugs or surgery.

There are four approaches:-

1) A normal gene is inserted to compensate for the dysfunctional gene.

2)An abnormal gene is traded for a normal gene. 3

3)An abnormal gene is repaired through selective reverse mutation.

4)Change the regulation of gene pair.
BRIEF HISTORY OF GENE THERAPY
US Office of
The National
Institute of Health Technology Jesse Gelsinger
Assessment
(NIH) took lead in stressed the becomes the
recombinant DNA difference first fatality in 637 GT clinical
(rDNA) research between somatic gene therapy. trials (3464
regulation. and germ-line patients)
therapy.

1974 1984 1999 2005

1967 1990 2003

1980
4

Nobelist The National FDA placed a

Marshall Dr. Martin J. Institute of temporary halt
Nirenberg Cline performs Health (NIH) on all gene
wrote for the first DNA performed therapy trials
programming transfer into the first using retroviral
cells. bone marrow approved vectors in
cells. gene therapy blood stem
procedure. cells.
 TYPES OF GENE THERAPY
Gene therapy is classified into two types:-
1) Somatic Gene Therapy
2) Germ-line Gene Therapy

1) SOMATIC GENE THERAPY

In somatic gene therapy, the desired
therapeutic gene is transferred to
somatic cells or stem cells of the
human body.
The gene is not transferred to the
offspring.
This is related to a single person and
the only person who has the
damaged cell will be replaced with 5
healthy cells.
This technique is considered the
best and safest method of gene
therapy.
Mechanism of Somatic gene therapy
2) GERM-LINE GENE THERAPY
In germ-line gene therapy, the
desired therapeutic gene is
introduced in the germ cells
(eggs or sperms) of the human
body.
The gene gets transferred from
one generation to another
generation.
This method is generally adopted
to treat the genetic, disease
causing-variations of genes that
are passed from the parents to
their children.
This therapy is not legal in many Mechanism of Germ-line gene therapy 6
places as the risk outweigh the
rewards.
 DIFFERENCES BETWEEN SOMATIC GENE
THERAPY AND GERM-LINE GENE THERAPY

In somatic gene therapy,

In germ-line gene therapy,
unhealthy cells are modified
with the correct gene and concerned genes are
transferred into gametic or
then transferred into the
patient’s body. early embryonic cells.

Cannot be inherited by
future generations.
Effective in target cells
Effective in all cells.
only.

Carried out multiple times Can be carried out once for

as it does not last for long. long lasting effects.

For safety, ethical and

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technical reasons, it is not
being attempted at present.

E.g., Introduction of genes

E.g., Genes introduced into
into bone marrow cells,
eggs and sperms.
blood cells, skin cells etc.
TYPES OF SOMATIC GENE THERAPY
SOMATIC GENE THERAPY

EX VIVO IN VIVO

Ex vivo gene therapy refers to

In vivo gene therapy refers to
the process of removing specific
the direct delivery of genetic
cells from a person, genetically
material either intravenously
altering them in a laboratory,
(through an IV) or locally to a
and then transplanting them
specific organ (e.g., directly into
back into the person.
the organ).
works by genetically
It works through the help of a
modifying a patient’s stem cells,
vector, which directly inserts
which then replace target cells
functional copies of a gene into
that have a missing or
target cells to treat a mutated
malfunctioning gene.
or missing gene.
This technique is used to treat 8
This technique has been proven
blood and immunological
in many areas of research.
diseases as well as genetic
Some of the currently approved
diseases that affect tissues and
gene therapies deliver genetic
organs that can be reached by
material in vivo.
blood cells.
 EX VIVO GENE THERAPY
Procedure for ex vivo gene therapy is as follows:-

Isolate cells with genetic defect

1 from a patient.

3 Introduce the therapeutic genes.

Select genetically corrected cells

4 and grow them. 9

Transplant the modified cells to

5 the patient.
 EX VIVO GENE THERAPY
Example:-
ADA (Adenosine deaminase) Deficiency
1st gene therapy(1990) – to correct the deficiency of
the enzyme, Adenosine deaminase (ADA).
It was performed on a 4-year-old girl Ashanthi
DeSilva.
She was suffering from SCID (Severe Combined
Immunodeficiency).
SCID Ashanti DeSilva

Caused due to a defect in the

gene coding for ADA.
Deoxyadenosine accumulates
and destroys T-lymphocytes.
Disrupts immunity, thus
patients suffer from
infectious diseases and die at 10
a young age.

Procedure to treat ADA deficiency

 IN VIVO GENE THERAPY
Procedure for ex vivo gene therapy is as follows:-

Therapeutic genes are inserted

1 in the vectors.

Genetically altered DNA is

2 injected into the patient’s body.

DNA is incorporated into the

3 target cells.

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Proteins encoded by the inserted

4 genes are produced.
 IN VIVO GENE THERAPY
Example:-
Cystic Fibrosis
Cystic fibrosis is a progressive,
genetic disease that causes
persistent lung infections and
limits the ability to breathe over
time.
In patients with cystic fibrosis, a
protein called Cystic Fibrosis
Transmembrane Regulator (CFTR)
is absent due to a gene defect.
In the absence of CFTR, chloride
ions concentrate within the cells
and draws water from the
surrounding.
This lead to the accumulation of
sticky mucous in the respiratory 12
tract and lungs.
Treated by in vivo replacement of
defective gene by adenovirus
vector.

Gene therapy for Cystic fibrosis

VECTORS IN GENE THERAPY
To transfer the desired gene into the target cell, a carrier or vehicle
is required. Such vehicles of gene delivery are known as ‘VECTORS’.
There are two main classes of vectors. Those are:-
1) Viral vectors
2) Non-viral vectors

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Viral vectors Non-viral vectors

IDEAL VECTORS
An ideal vector must have the following
features:-
Target the right cells.
Small size and low molecular weight.
Integrate the gene in the cells.
Activate the gene.
Avoid harmful side effects
In the real world, no universal vector exists.

1) VIRAL VECTORS
Viruses introduce their genetic material into
the host cell as part of their replication cycle.
These are used as vectors by removing their
viral DNA and inserting the therapeutic DNA 14
in them.
The viruses used are altered to make them
safe, although some risks still exist with this
technique of gene therapy.
TYPES OF VIRAL VECTOR
A number of viruses have been used for human gene therapy including :
1) Retrovirus
2) Adenovirus
3) Adeno-associated virus
4) Herpes simplex virus
1) RETROVIRUS
The recombinant retroviruses have the ability to
integrate into the host genome.
The typical maximum length of an allowable DNA
insert in a retroviral vector is about 8-10 kB.
Target – actively dividing cell.

Retrovirus
15

Action of Retrovirus
 a) LENTIVIRUS
Subclass of retroviruses.
The viral genome in the form of RNA is reverse-
transcribed when the virus enters the cell to produce
DNA, which is then inserted into the genome at a random
position via the viral integrase enzyme.
Target – dividing, non-dividing cells Lentivirus

2) ADENOVIRUS
Adenoviral DNA does not integrate into the genome and
is not replicated during cell division.
Humans commonly come in contact with adenovirus. The
majority of patients have already developed neutralizing
antibodies which can inactivate the virus.
Target – dividing, non-dividing cells
3) ADENO-ASSOCIATED VIRUS Adenovirus

It is a small virus and it affects humans and other primates.

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It enters the host cell, becomes double-stranded, and gets
integrated into chromosomes.
It is not currently known to cause disease and consequently
the virus causes a very mild immune response.
Target – non-dividing, dividing cells
Adeno- associated virus
 4) HERPES SIMPLEX VIRUS
The herpes simplex virus is a human neurotropic
virus.
This is mostly examined for gene transfer in the
nervous system.
These viruses have a natural tendency to infect a
particular type of cell. Herpes simplex virus

ADVANTAGES AND DISADVANTAGES OF USING VIRAL VECTORS

They can cause immune

amount of genetic
materials. Therefore some
genes may be too big to fit
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into some viruses.

DISADVANTAGES
2) NON- VIRAL VECTORS SYSTEM

1) PURE DNA EXTRACT

Direct introduction of pure DNA
construct into the target tissue.
Efficiency of DNA uptake by cells and
expression is rather low.
Consequently, large quantities of DNA
have to be injected periodically.

Pure DNA extract

2) DNA MOLECULAR CONJUGATES
Commonly used synthetic conjugate is
poly-L-lysine bound to specific target
cell receptors.
Therapeutic DNA is then made to 18
combine with the conjugate to form a
complex.
It avoids the lysosomal breakdown of
DNA.
Poly-L-lysine
 3) LIPOPLEXES
Lipid DNA complexes.
DNA construct surrounded by artificial lipid
layer.
Most of it gets degraded by lysosomes.

4) HUMAN ARTIFICIAL CHROMOSOME

Can carry a large DNA i.e., with one or
more therapeutic genes.

ADVANTAGES AND DISADVANTAGES OF USING NON-VIRAL VECTORS

ADVANTAGES

Easy to produce
19
Good transduction of some
cell types
There is no limit to the
transgene size DISADVANTAGES
Useful for local therapy
METHODS OF GENE DELIVERY
1) PHYSICAL METHODS
i) GENE GUN/ BIOLISTICS
It uses a gun-like instrument to send
desired genes inside host cells.
It introduces the desired rDNA into a
plant cell by coating it with gold or
tungsten and firing it into the tissue at
high velocity.
ii) MICROINJECTION Gene gun Mechanism of gene gun

In this method, the host cells are forced to

take up the desired rDNA molecules using
microinjection.
20
This is a physical method that requires the
usage of a specialized optical microscope.
The host cell is selected, and the rDNA is
injected into the host cell.
This method is generally used to introduce
rDNA into animal cells. Microinjection
2) CHEMICAL METHODS
i) USING DETERGENT MIXTURE
Certain charged chemical compounds
like calcium phosphates are mixed with
functional cDNA of the desired
function.
The mixture is introduced near the
vicinity of recipient cells.
The chemicals disturb the cell Gene transfer using detergent mixture
membrane, widen the pore size, and
allow cDNA to pass through the cells.

ii) LIPOFECTION
It is a technique used to inject genetic
materials into a cell using
transfection agents like liposomes.
21
Liposomes are artificial phospholipid
vesicles used to deliver a variety of
molecules including DNA into the
cells.
Mechanism of lipofection
SUCCESS CASES OF GENE THERAPY
1) SICKLE CELL ANEMIA
It is a genetic condition that affects the
RBCs.
It affects the hemoglobin in the blood
and causes RBCs to be rigid and shaped
like the letter ‘C’ or a sickle
Until recently, a bone marrow transplant
is the only cure for sickle cell anemia.
Gene therapy for Sickle cell anemia
2) PARKINSON DISEASE
It is a long-term degenerative disorder of the
central nervous system that mainly affects
the motor system.
Gene therapy in Parkinson’s disease consists
of the creation of new cells that produce a 22
specific neurotransmitter (dopamine),
protect the neural system, or modification of
genes that are related to the disease.

Gene therapy for Parkinson disease

PROBLEMS WITH GENE THERAPY
SHORT-LIVED NATURE OF GENE THERAPY:
1 Patients will have to undergo multiple rounds of gene therapy.

IMMUNE RESPONSE:
Stimulates the immune system that reduces gene therapy
effectiveness.
2

PROBLEMS WITH VIRAL VECTORS:

3 Potential problems to the patient like toxicity, immune and
inflammatory responses, and gene control targeting are created.

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MULTI GENE DISORDER :
Disorders like heart disease, high blood pressure, arthritis, and
diabetes are caused by the combined effect of many genes.
4
ETHICAL ISSUES OF GENE THERAPY
Is it all right to use
What is normal the therapy in the
and what is the Should people be prenatal stage of
disability or allowed to use gene development in
disorder and who therapy to enhance babies?
decides? basic human traits such
as height, intelligence,
etc.? 5
1
3 Preliminary attempts at
Who will have gene therapy are
access to your expensive. Who will have
genetic Is it interfering access to these therapies?
information? with God’s plan? Who will pay for their use?
24

2 4
6
 CONCLUSION
Theoretically, gene therapy is the permanent solution
1
g for genetic diseases. But some treatments are short-
lived and have to be repeated.

2 But it has several complexities. At its current stage, it

I is not accessible to most people due to its huge cost.

3 A breakthrough may come at any time, and a day may come

when almost every disease will have gene therapy.

4
Gene therapy has the potential to revolutionize the 25
practice of medicine.
 BIBLIOGRAPHY
NCERT Biology Book – XII
www.youtube.com
www.googleimages.com
http://en.wikipedia.org/wiki/Gene_ therapy
http://en.wikipedia.org/wiki/Somatic_ gene_ therapy
http://en.wikipedia.org/wiki/Germ_ line_ gene_ therapy
http://www.medindia.net/articles/genetherapy_ treatment.htm
www.byjus.com
And more…

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THANK YOU!