APPENDIX 1

STUDY ON GENE THERAPY

BIOLOGY PROJECT REPORT

by

ANURAG JOSHUA
04 BiPC

DEPARTMENT OF BIOLOGY
MOUNT CARMEL GLOBAL SCHOOL
HYDERABAD
2023-2024
 APPENDIX 2

BONAFIDE CERTIFICATE

This is to certify that this project report entitled “STUDY ON GENE

THERAPY”, submitted to MOUNT CARMEL GLOBAL SCHOOL,
HYDERABAD, is a bonafide record of work done by “ANURAG
JOSHUA” under my supervision
from “………………..” to “………………..”

Signature

Mrs DHANASHRI SAMUDRALWAR

(BIOLOGY TEACHER)

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 APPENDIX 3

Declaration by Author

This is to declare that this report has been written by me. No part of the
report is plagiarized from other sources. All information included from other
sources have been duly acknowledged. I aver that if any part of the report is
found to be plagiarized, I shall take full responsibility for it.

Signature

Anurag Joshua
04

Place
Date

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 APPENDIX 4
(A typical specimen of table of contents)

TABLE OF CONTENTS

1. ABSTRACT

2. INTRODUCTION

3. HISTORY OF GENE THERAPY

4. TYPES OF GENE THERAPY

5. SOMATIC GENE THERAPY

6. GERM LINE GENE THERAPY

7. VIRAL VECTORS

8. OUTCOMES OF GENE THERAPY

9. FUNCTIONAL CLASSIFICATION

10. GENETIC DISORDERS

- TARGETS FOR GENE THERAPY

- TECHNIQUES USED IN GENE THERAPY

11. GENE TARGETING

12. SUCCESS OF GENE THERAPY

13. ADVANTAGES AND DISADVANTAGES

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14. CHALLENGES

15. RECENT UPCOMING

16. CONCLUSION

17. REFERENCES

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 ABSTRACT

Gene therapy is the introduction of genes into existing cells to prevent or

cure a wide range of diseases. It is a technique for correcting defective genes
responsible for disease development. The first approved gene therapy
experiment occurred on September 14, 1990 in US when Ashanti DeSilva
was treated for ADA-SCID. Gene therapy is designed to introduce genetic
material into cells to compensate for abnormal genes or to make a beneficial
protein. If a mutated gene causes a necessary protein to be faulty or missing,
gene therapy may be able to introduce a normal copy of the gene to restore
the function of the protein. A gene that is inserted directly into a cell usually
does not function. Instead, a carrier called a vector is genetically engineered
to deliver the gene. Certain viruses are often used as vectors because they
can deliver the new gene by infecting the cell. The viruses are modified so
they can't cause disease when used in people. Some types of virus, such as
retroviruses, integrate their genetic material (including the new gene) into a
chromosome in the human cell. Other viruses, such as adenoviruses,
introduce their DNA into the nucleus of the cell, but the DNA is not
integrated into a chromosome.

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 INTRODUCTION

Gene therapy is an experimental technique that uses genes to treat or prevent

disease. In the future, this technique may allow doctors to treat a disorder
by inserting a gene into a patient’s cells instead of using drugs or surgery.
Researchers are testing several approaches to gene therapy, including:
 Replacing a mutated gene that causes disease with a healthy copy of
the gene.
 Inactivating or “knocking out” a mutated gene that is functioning
improperly.
 Introducing a new gene into the body to help fight a disease.
Although gene therapy is a promising treatment option for a number of
diseases (including inherited disorders, some types of cancer, and certain
viral infections), the technique remains risky and is still under study to
make sure that it will be safe and effective. Gene therapy is currently being
tested only for diseases that have no other cures. An experimental
technique for correcting defective genes that are responsible for disease
development.
The most common form of gene therapy involves a normal gene to replace
an abnormal gene. It is intracellular delivery of genes to generate a
therapeutic effect by correcting an existing abnormality.

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 HISTORY AND DEVELOPMENT OF GENE THERAPY

1960: The concepts of gene therapy was introduced.

1970: Friedmann and Roblin author of paper in science titled “Gene therapy
for human genetic disease?” cite the first attempt to perform gene therapy.
1990: The first approved gene therapy case at the National Institute of
Health, U.K. It was performed on four year old girl named Ashanti DaSilva.
It was a treatment for a genetic defect that left her with an immune system
deficiency.
Now gene therapy approach repairs errors in messenger RNA derived from
defective gene. This technique has the potential to treat the blood disorder
Thalassaemia, Cystic fibrosis, and some cancers.
Sickle cell disease is successfully treated in mice.
1992: Doctor Claudio Bordignon working at the Vita-Salute San Raffaele
University, Milan, Italy, performed the first procedure of gene therapy using
hematopoietic stem cells as vectors to deliver gene intended to correct
hereditary diseases.
1999: Death of Jesse Gelsinger in a gene therapy experiment resulted in a
significant setback to gene therapy research in the United States.
2006: Scientists at the National Institute of Health (Bethesda, Maryland)
have successfully treated metastatic melanoma in two patients. This study
constitutes one of the first demonstrations that gene therapy can be effective
in treating cancer.
2007-2011: Research is still ongoing and the number of diseases that has
been treated successfully by gene therapy increases.
 Retinal disease
 Colour blindness
 Adrenoleukodystrophy
2011: Medical community accepted that it can cure HIV as in 2008, Gero
Hutter has cured a man from HIV using gene therapy.

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 TYPES OF GENE THERAPY

1. Germ line gene therapy

2. Somatic gene therapy

GERM LINE GENE THERAPY

Therapeutic genes transferred into the germ cells. It is heritable and passed
on to later generations. Result in permanent changes. Potential for offering
a permanent therapeutic effect for all who inherit the target gene. Possibility
of eliminating some diseases from a particular family.
E.g. Genes introduced into eggs and sperms.

SOMATIC CELL GENE THERAPY

Therapeutic genes transferred into the somatic cells. Affects only the target
cells in the patient, and is not passed to future generations. Short-lived
because the cells of most tissues ultimately die and are replaced by new
cells. Transporting the gene to the target cells or tissue is also problematic.
E.g. Introduction of genes into bone marrow cells, blood cells, skin cells,
etc.

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 VIRAL VECTORS

In both types of therapy, scientists need something to transport either the

entire gene or a recombinant DNA to the cell's nucleus, where the
chromosomes and DNA reside. In essence, vectors are molecular delivery
trucks. One of the first and most popular vectors developed were viruses
because they invade cells as part of the natural infection process. Viruses
have the potential to be excellent vectors because they have a specific
relationship with the host in that they colonize certain cell types and tissues
in specific organs. Asa result, vectors are chosen according to their attraction
to certain cells and areas of the body.
One of the first vectors used was retroviruses. Because these viruses are
easily cloned (artificially reproduced) in the laboratory, scientists have
studied them extensively and learned a great deal about their biological
action. They also have learned how to remove the genetic information that
governs viral replication, thus reducing the chances of infection.
Retroviruses work best in actively dividing cells, but cells in the body are
relatively stable and do not divide often. As a result, these cells are used
primarily for ex vivo (outside the body) manipulation. First, the cells are
removed from the patient's body, and the virus, or vector, carrying the gene
is inserted into them. Next, the cells are placed into a nutrient culture where
they grow and replicate. Once enough cells are gathered, they are returned
to the body, usually by injection into the blood stream. Theoretically, as long
as these cells survive, they will provide the desired therapy.
Another class of viruses, called the adenoviruses, also may prove to be good
gene vectors. These viruses can effectively infect non-dividing cells in the
body, where the desired gene product then is expressed naturally. In addition
to being a more efficient approach to gene transportation, these viruses,
which cause respiratory infections, are more easily purified and made stable
than retroviruses, resulting in less chance of an unwanted viral infection.
However, these viruses live for several days in the body, and some concern
surrounds the possibility of infecting others with the viruses through
sneezing or coughing. Other viral vectors include influenza viruses, Sindbis
virus, and a herpes virus that infects nerve cells.
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Scientists also have delved into nonviral vectors. These vectors rely on the
natural biological process in which cells uptake (or gather) macromolecules.
One approach is to use liposomes, globules of fat produced by the body and
taken up by cells. Scientists also are investigating the introduction of raw
recombinant DNA by injecting it into the bloodstream or placing it
on microscopic beads of gold shot into the skin with a "gene-gun." Another
possible vector underdevelopment is based on dendrimer molecules. A class
of polymers (naturally occurring or artificial substances that have a high
molecular weight and formed by smaller molecules of the same or similar
substances), is "constructed" in the laboratory by combining these smaller
molecules. They have been used in manufacturing Styrofoam, polyethylene
cartons, and Plexiglas. In the laboratory, dendrimers have shown the ability
to transport genetic material into human cells. They also can be designed to
form an affinity for particular cell membranes by attaching to certain sugars
and protein groups.

OUTCOMES OF GENE THRAPY

• Replaces a mutated gene with a healthy one.

• Deactivates the gene that isn’t functioning properly.

• Introduces a new gene in the body to help fight the disease.

• Enhances the affect of a normally functioning gene.

• Activates the gene that was shut down during fetal life.

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 FUNCTIONAL CLASSIFICATION

Based on the purpose of gene therapy, it can be classified as –

1. Gene replacement therapy
2. Gene deactivation therapy
3. Transgenesis
4. Gene Enhancement therapy
5. Gene activation therapy

GENETIC DISORDERS

A genetic disorder is an illness caused by one or more abnormalities in the

genome, especially a condition that is present from birth (congenital). They
are medical disorders related to gene mutation. Genetic disorders are
heritable, and are passed down from the parents’ genes. Other defects may
be caused by new mutations or changes to the DNA. In such cases, the defect
will only be heritable if it occurs in the germ line. The same disease, such
as some forms of cancer, may be caused by an inherited genetic condition
in some people, by new mutations in other people, and by non-genetic
causes in still other people. These diseases are totally random and difficult
to prevent as they are not caused by external agents. Also, as their root
causes lies in the genome of the organism their cure was thought to be
impossible until the breakthrough research unlocking the secrets of DNA
leading to the development of biotechnology and hence gene therapy.
We can think of a medical condition or illness as a “broken window”. Many
medical conditions result from flaws, or mutations, in one or more of a
person’s gene. Mutations cause the protein encoded by that gene to
malfunction. When a protein malfunctions, cells that rely on that protein’s
function can’t behave normally, causing problems for whole tissues or
organs. Medical conditions related to gene mutations are called genetic
disorders.

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So, if a flawed gene caused our “broken window”, can we “fix” it? What are
our options?
1. Stay silent: ignore the genetic disorder and nothing gets fixed.

2. Try to treat the disorder with drugs or other approaches: depending on

the disorder, treatment may or may not be a good long-term situation.

3. Put in a normal, functional copy of the gene: if you can do this, it may
solve the problem!

If it is successful, gene therapy provides a way to fix a problem at its source.

Adding a corrected copy of the gene may help the affected cells, tissues and
organs work properly. Gene therapy differs from traditional drug-based
approaches, which may treat the problem, but which do not repair the
underlying genetic flaw.

TARGETS FOR GENE THERAPY

But now a question arises, which disorders or diseases can we target using
gene therapy? Many disorders or medical conditions might be treated using
gene therapy, but others may not be suitable for this approach. For a disease
to be targeted by gene therapy it must satisfy the following conditions:
1. The condition must result from mutations in one or more genes

2. To treat a genetic flaw, the knowledge of which gene(s) to pursue is

absolutely necessary. Also a DNA copy of that gene available in the
laboratory. The best candidates for gene therapy are the so-called
“single-gene” disorders – which are caused by mutations in only one
gene.

3. To design the best possible approach, knowledge about how the gene
factors into the disorder is required.

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For example:
 Which tissues are affected?
 What roles foes the protein encoded by the gene play within the cells
of that tissue?
 Exactly how do mutations in the gene affect the protein’s function?

4. Adding a normal copy of the gene should fix the problem in the
affected tissue. This may seem like obvious, but it’s not. What if the
mutated gene encodes a protein that prevents the normal protein from
doing it’s job? Mutated genes that function this way are called
dominant negative and adding back the normal protein won’t fix the
problem.

5. The gene delivery to cells of the affected tissue must be possible. It

depends on:

 How accessible is the tissue? Is it fairly easy (skin, blood or lungs),

or more difficult to reach (internal organs)?

 What is the best mode of delivery?

TECHINIQUES USED IN GENE THERAPY

The techniques of biotechnology have made it possible to isolate the

required gene in the laboratory and also deliver the gene.

Isolation of Gene of Interest

The first step is to find and isolate the gene that will be inserted into the
genetically modified organism. Finding the right gene to insert usually
draws on years of scientific research into the identity and function of
useful genes. Once that is known the DNA needs to be cut at specific
locations to isolate the gene of interest. This can be done by using

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restriction enzyme also known as molecular scissors which cut DNA at
specific sites containing palindromic DNA sequences. But in order to cut
the DNA with restriction enzymes, it needs to be in pure form, free from
other macro-molecules.

Isolation of DNA

Since the DNA is enclosed within the membranes, we have to break the
cell open to release DNA along with other macromolecules such as RNA,
proteins, polysaccharides and also lipids. This can be achieved by
treating the bacterial cells/plant or animal tissue with enzymes such as
lysozyme (bacteria), cellulase (plant cells), chitinase (fungus). Genes are
located on long molecules of DNA intertwined with proteins such as
histones. The RNA can be removed by treatment with ribonuclease
whereas proteins can be removed by treatment with protease. Other
molecules can be removed by appropriate treatments and purified DNA
ultimately precipitates out after the addition of chilled ethanol. This can
be seen as collection of fine threads in the suspension.

Cutting of DNA

Restriction enzymes digestions are performed by incubating purified

DNA molecules with the restriction enzyme, at the optimal conditions or
that specific enzyme. The cutting of DNA by restriction endonucleases
results in the fragments of DNA. These fragments can be separated by a
technique known as gel electrophoresis. Since DNA fragments are
negatively charged molecules, they can be separated by forcing them to
move towards the anode under an electric field through a medium/matrix.
The separated bands of DNA are analysed for the required gene and then
it is cut out from the agarose gel and extracted from the gel piece. This
step is known as elution.

Multiplication of Gene (PCR)

PCR or polymerase chain reaction is then used to create multiple copies

of the gene of interest. In this reaction, multiple copies of the gene (or

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DNA) of interest is synthesised in vitro using two sets of primers (small
chemically synthesised oligonucleotides that are complementary to the
regions of DNA) and the enzyme DNA polymerase. The enzyme extends
the primers using the nucleotides provided in the reaction and the
genomic DNA as template. If the process of replication of DNA is
repeated many times, the segment of DNA can be amplified to
approximately billion times, i.e., 1 billion copies are made.

GENE TARGETING

Gene delivery is one of the biggest challenges in the field of gene therapy.

Gene delivery includes:

1. TARGETTING the right cells.

2. ACTIVATING the gene. A gene’s journey is not over when it

enters the cells. It must go to the cell’s nucleus and be “turned on”,
meaning that its transcription and translation are activated to
produce the protein product encoded by the gene. For gene delivery
to be successful, the protein that is produced must function
properly.

3. INTEGRATING the gene in the cells. The gene must stay put and
continue working in the target cells. If so, it must be ensured that
the gene integrates into, or becomes part of the host cell’s genetic
material, or that the gene finds another way to survive in the
nucleus without being rejected.

4. AVOIDING harmful side effects. Anytime an unfamiliar

biological substance is introduced into the body, there is a risk that
it will be toxic or that the body will mount an immune response
against it. If the body develops immunity against a specific gene
delivery vehicle, future rounds of the therapy will be ineffective.

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Choosing The Best Vector

There is no “perfect vector” that can treat every disorder. Like any type of
medical treatment, a gene therapy vector must be customized to address the
unique features of the disorder. We have learnt the lesson, of transferring
genes into plants and animals from bacteria and viruses, which have known
this for ages - how to deliver genes to transform eukaryotic cells and force
them to do what the bacteria or viruses want.
Part of the challenge in gene therapy is choosing the most suitable vector
for treating the disorder. Some vectors commonly used are:

 Viruses

Usually when we think of viruses, we think of them causing diseases

such as the common cold, the flu, and HIV/AIDS. When faced with the
problem of genetic delivery, scientists looked to viruses. Why reinvent
the wheel if there’s a perfectly good one out there? If we can modify
viruses to deliver genes without making people sick, we may have a good
set of gene therapy tools.

General Advantages of Viral Vectors:

 They’re very good at targeting and entering cells.

-Some viral vectors might be engineered to target specific types of

cells.

-They can be modified so that they can’t replicate and destroy the
cell.

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General Drawbacks of Viral Vectors:

A virus can’t “expand” to fit a piece of genetic material larger than it is

naturally built to carry. Therefore, some genes may be too big to fit into a
certain type of virus.

 Viruses can cause immune responses in patients, resulting in two

potential outcomes:
- Patients may get sick.

 A patient’s immunity to a virus may prevent him from responding

to repeated treatments.

 However, modern viral vectors have been engineered without

most of the proteins that would cause an immune response.

Non-Viral Vectors

 Although viruses can effectively deliver genetic material into a

patient’s cells, they do have some limitations. It is sometimes more
efficient to deliver a gene using a non-viral vector, which has fewer
size constraints and which won’t generate an immune response.
 Non-viral vectors are typically circular DNA molecules, also known
as plasmids. In nature, bacteria use plasmids to transfer genes from
cell to cell.

Scientists use bacteria and plasmids to easily and efficiently store and
replicate genes of interest from any organism.
Vectors used at present, are engineered is such a way that they help easy
linking of foreign DNA and selection of recombinants from non-
recombinants.

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These are not the only way to introduce alien DNA into host cells. In a
method known as micro-injection, recombinant DNA is directly
injected into the nucleus of an animal cell. In another method, suitable
for plants, cells are bombarded with high velocity micro-particles of
golf or tungsten coated with DNA in a method known as biolistics or
gene gun.

Cystic Fibrosis
Cystic Fibrosis (CF), also known as mucoviscidosis, is an autosomal
recessive genetic disorder that affects most critically the lungs, and also
the pancreas, liver, and intestine. It is characterised by abnormal
transport of chloride and sodium across an epithelium, leading to thick,
viscous secretions, preventing the cilia from clearing debris which
cause symptoms such as coughing, poor digestion and increased
vulnerability to infection.
CF is caused by a mutation in the gene for the protein cystic fibrosis
transmembrane conductance regulator (CFTR) gene on chromosome 7.
Most commonly, the mutation in the CFTR gene is a three-base-pair
deletion. This protein is required to regulate the components of sweat,
digestive fluids, and mucus. CFTR regulates the movement of chloride
and sodium ions across epithelial membranes, such as the alveolar
epithelial located in the lungs. Since all of the cells of a CF patient have
the defective protein, large quantities of thick, sticky mucus build up
throughout the lungs and other organs. This results in the severity of
symptoms seen in CF patients.

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 SUCCESS CASES OF GENE THERAPY

1. Gene Therapy Cures Blindness

Cure blindness of inherited condition.
Leber’s congenital amaurosis
- Inherited disease caused by an abnormality in a gene called
RPE65.
- This condition appears at birth or in the first few months of life
and causes progressive worse and loss of vision.

How It Works?
 Used harmless viruses
 Enable access to the cells beneath the retinas of patients.
 By using a very fine needle – safe in an extremely fragile tissue and
can improve vision in a condition previously considered wholly
untreatable.

2. Gene Therapy Reduces Parkinson’s Disease Symptoms

 It significantly improved the weakness of the symptoms
such as tremors, motor skill problems, and rigidity.
 Main- overactive brain region: the subthalamic nucleus
should be introduced with gene.
 That would produce GABA- an inhibitory chemical- then
they could potentially quiet that brain region and alleviate
tremors.

How It Works?
 Done with local anesthesia, used a harmless, inactive virus [AAV-2
GAD]
 Deliver the GAD gene into the patient’s subthalamic nucleus.

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  The gene instructs cells to begin making GABA neurotransmitters to
re-establish the normal chemical balance that becomes dysfunctional
as the diseases progresses.

ADVANTAGES

 Give a chance of normal life to baby born with genetic disease.

 Give hope of healthy life to cancer patients.
 For certain disease that do not have any cure except gene therapy, it
could save many lives.

DISADVANTAGES

 The genetic testing, screening and research in finding the availability

of certain gene is very controversial.
 May increase the rate of abortion if parental test regarding baby with
genetic disease is done.
 The cost is very high and the patient might need an insurance to
cover the treatment.
 Cosmetic industry may monopolized this gene therapy if it is used in
enhancing beauty and in vanishing the aging effect, rather than used
for treatment of a disease.

Is It A Good Target For Gene Therapy?

To check this, some questions must be answered:
1. Does the condition result from mutation?
Yes.
2. Is the biology of the disorder known?
Yes.
3. Will adding a normal copy of the gene fix the problem in the
affected tissue?
Yes. While the mutated CFTR gene encodes a non-functional ion
channel protein, it will not prevent a normal CFTR channel protein
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from working properly. Therefore, adding a normal copy of the CFTR
gene should fix the problem.
Is it feasible to deliver the gene to the cells of the affected tissue? Yes,
in part. Treating the lungs of patients with CF might be feasible, since
the lung surfaces are exposed to the air and somewhat easy to reach.
Because the digestive system is less accessible, however, it might be a
more difficult region to treat.
Hence, we can conclude that it is a perfected disease to be treated by
gene therapy.

CHALLENGES

Some of the factors that have kept gene therapy from becoming an
effective treatment for genetic diseases are:

 Short- lived nature of gene therapy -

Before gene therapy can become permanent cure for any condition, the
therapeutic DNA introduced into the target cells must remain functional
and the cells containing the therapeutic DNA must be long-lived and
stable. Problems with integrating therapeutic DNA into the genome and
the rapidly dividing nature of many cells prevent gene therapy from
achieving any long-term benefits. Patients will have to undergo multiple
rounds of gene therapy.

 Immune response -
Anytime a foreign body is introduced into human tissues, the immune
system is designed to attack the invader. The risk of stimulating the
immune system in a way that reduces gene therapy effectiveness is
always a potential risk. Furthermore, the immune system enhanced
response to invaders it has seen before makes it difficult for gene therapy
to be repeated in patients.

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 Problems with viral vectors -
Viruses, while the carrier of choice in most gene therapy studies, present
a variety of potential problems to the patient – toxicity, immune and
inflammatory responses, and gene control and targeting issues. In
addition, there is always the fear that the viral vector, once inside the
patient, may recover its ability to cause disease.

 Multigene disorders -
Conditions or disorders that arise from mutations in the single gene are
the best candidates for gene therapy. Unfortunately, some of the most
commonly occurring disorders, such as heart disease, high blood
pressure, Alzheimer’s disease, arthritis, and diabetes are caused by the
combined effects of variations in many genes. Multigene or
multifractional disorders such as these would be especially difficult to
treat effectively using gene therapy.

RECENT UPCOMING

CRISPR
CRISPR stands for Clustered Regularly Interspaced Short Palindromic
Repeats. These RNA sequences serve an immune function in
archaebacteria, but in the last year or so, scientists have seized upon them
to rewrite genes. The RNA sequence serves as a guide to target a DNA
sequence in, say, a zygote or a stem cell. The guide sequence leads an
enzyme, Cas9, to the DNA of interest. Cas9 can cut the double strand,
nick it, or even knock down gene expression. After Cas9 injuries the
DNA, repair systems fix the sequence or new sequences can be inserted.

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It isn’t the first or only method of gene repair therapy that’s been
developed, but the CRISPR technology, says Ramesar, is so special
because, unlike previous methods which were more laborious and could
only target one kind of cell in the body, it appears to be a “one size fits
all delivery”, adaptable for different tissues. The procedure also seems
relatively simple to perform.
Exciting as the development may be, CRISPR won’t be delivering instant
cures just yet.
Ramesar says, from his initial impressions of the literature, that it would
seem that localised, accessible abnormal tissue (as in the retina or skin)
could be targeted more easily.
Conditions affecting the body more systemically, however, such as
certain developmental syndromes, or central nervous system disorders,
might be problematic in terms of getting the repair technology into
enough of the target cells in that tissue to make an effective difference.

CONCLUSION

Although early clinical failures led many to dismiss gene therapy as over-
hyped, clinical successes since 2006 have bolstered new optimism in the
promise of gene therapy. These include successful treatment of patients
with the retinal disease Leber’s congenital amaurosis, X-linked SCID,
ADA-SCID, adrenoleukodystrophy, chronic lymphocytic leukaemia
(CLL), acute lymphocytic leukaemia (ALL), multiple myeloma,
haemophilia and Parkinson’s disease. These recent clinical success have
led to a renewed interest in gene therapy, with several articles in scientific
and popular publications calling for continued investment in the field.

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 REFERENCES

1. What is gene therapy https://www.nhlbi.nih.gov/health/genetic-

therapies
2. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2907101/

3. Gene therapy information https://www.uniqure.com/assets/uploads/Gene-Therapy-

Information.pdf
4. https://en.wikipedia.org/wiki/Gene_therapy
5. Gene therapy treatment
https://learn.genetics.utah.edu/
6. https://www.genome.gov/genetics-glossary/Gene-
Therapy#:~:text=Gene%20therapy%20is%20a%20technique,healthy
%20version%20of%20that%20gene.
7. Handbook on therapy https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1051858/
8. Cystic Fibrosis https://www.cff.org/research-clinical-trials/gene-therapy-cystic-fibrosis
9. https://www.wikipedia.org/
10.https://www.yourgenome.org/facts/what-is-gene-therapy/
11.https://knowgenetics.org/gene-therapy/
12.https://www.sciencedaily.com/terms/gene_therapy.htm

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