Certificate

This is To cerTify ThaT This “biology invesTigaTory

projecT” on The Topic “gene Therapy” has been
successfully completed by Prasun Saren of class 12th
(science) under the guidance of Mr. jagjit patel(pgt
biology) in particular fulfilment of the curriculum of
central board of secondary education(cbse) leading
to the award of annual examination of the year 2019-
2020

TEACHER-IN-CHARGE EXAMINER
 ACTKNOWLEDGEMENT
I WOULD LIKE TO express my special
thanks of gratitude to my teacher Mr. Jagjit
Patel, who gave me the golden opportunity to
do this wonderful project of biology on “gene
therapy”, who also helped me in completing my
project. I came to about so many new things I
am really thankful to them. Secondly I would
also like to thank my friends who helped me in
finalizing this project within the time frame.

12th Science
Prasun Saren
Topic
 INTRODUCTION
What is GENE THERAPY?
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.

Gene therapy (also called human gene transfer) is the therapeutic delivery of nucleic
acid into a patient's cells as a drug to treat disease.

BACKGROUND:-
Gene therapy was conceptualized in 1972, by authors who urged caution before
commencing human gene therapy studies.
The first attempt, an unsuccessful one, at gene therapy (as well as the first case of
medical transfer of foreign genes into humans not counting organ transplantation) was
performed by Martin Cline on 10 July 1980. Cline claimed that one of the genes in his
patients was active six months later, though he never published this data or had it
verified and even if he is correct, it's unlikely it produced any significant beneficial
effects treating beta-thalassemia.
After extensive research on animals throughout the 1980s and a 1989 bacterial gene
tagging trial on humans, the first gene therapy widely accepted as a success was
demonstrated in a trial that started on 14 September 1990, when Ashi DeSilva was
treated for ADA-SCID
The first somatic treatment that produced a permanent genetic change was initiated in
1993. The goal was to cure malignant brain tumors by using recombinant DNA to
transfer a gene making the tumor cells sensitive to a drug that in turn would cause the
tumor cells to die.
Gene therapy is a way to fix a genetic problem at its source. The polymers are
either translated into proteins, interfere with target gene expression, or possibly
correct genetic mutations.
The most common form uses DNA that encodes a functional, therapeutic gene to
replace a mutated gene. The polymer molecule is packaged within a "vector", which
carries the molecule inside cells.

TYPES OF GENE THERAPY

SOMATIC GERMLINE

SOMATIC:-
In somatic cell gene therapy (SCGT), the therapeutic genes are transferred into any cell
other than a gamete, germ cell, gametocyte, or undifferentiated stem cell. Any such
modifications affect the individual patient only, and are not inherited by offspring.
Somatic gene therapy represents mainstream basic and clinical research, in which
therapeutic DNA (either integrated in the genome or as an external episome or plasmid)
is used to treat disease.
Over 600 clinical trials utilizing SCGT are underway in the US. Most focus on severe
genetic disorders, including immunodeficiencies, haemophilia, thalassaemia, and cystic
fibrosis. Such single gene disorders are good candidates for somatic cell therapy. The
complete correction of a genetic disorder or the replacement of multiple genes is not yet
possible. Only a few of the trials are in the advanced stages.
GERMLINE:-
In germline gene therapy (GGT), germ cells (sperm or egg cells) are modified by the
introduction of functional genes into their genomes. Modifying a germ cell causes all the
organism's cells to contain the modified gene. The change is therefore heritable and
passed on to later generations. Australia, Canada, Germany, Israel, Switzerland, and
the Netherlands prohibit GGT for application in human beings, for technical and ethical
reasons, including insufficient knowledge about possible risks to future generations and
higher risks versus SCGT. The US has no federal controls specifically addressing
human genetic modification (beyond FDA regulations for therapies in general).

How does gene therapy work?

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.
The vector can be injected or given intravenously (by IV) directly into a specific tissue in
the body, where it is taken up by individual cells. Alternately, a sample of the patient's
cells can be removed and exposed to the vector in a laboratory setting. The cells
containing the vector are then returned to the patient. If the treatment is successful, the
new gene delivered by the vector will make a functioning protein.

Researchers must overcome many technical challenges before gene therapy will be a
practical approach to treating disease. For example, scientists must find better ways to
deliver genes and target them to particular cells. They must also ensure that new genes
are precisely controlled by the body.

A new gene is inserted directly into a cell. A carrier called a vector is genetically
engineered to deliver the gene. An adenovirus introduces the DNA into the nucleus of
the cell, but the DNA is not integrated into a chromosome.

Gene Targeting:-
Gene delivery is one of the biggest challenges in the field of gene therapy.
Gene Delivery includes:
1. TARGETING the right cells.
2. ACTIVATING the gene. A gene's journey is not over when it enters the cell. 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.

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 gene 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.
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 in such a way that they help easy linking of
foreign DNA and selection of recombinants from non-recombinants.
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 gold or tungsten coated with DNA in a method known as biolistic or gene gun.

CHALLENGES:-
Some 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 a permanent cure for any condition, the therapeutic
DNA introduced into 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 object 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's
enhanced response to invaders it has seen before makes it difficult for gene therapy to
be repeated in patients.
• 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 a single gene are the best candidates
for gene therapy. Unfortunately, some 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 multifactorial
disorders such as these would be especially difficult to treat effectively using gene
therapy.

CONCLUSIONS
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 successes have led to a renewed interest in gene therapy, with several articles in
scientific and popular publications calling for continued investment in the field.

BIBLIOGRAPY:-
 WIKIPEDIA
 SCIENCE DAILY