Gene Manipulation

A PROJECT REPORT
SUBMITTED TO
NATIONAL MODEL SENIOR SECONDARY SCHOOL
AFFILIATED TO CBSE FOR THE AISSCE 2025

BY
S.I.VISHALI
UNDER THE GUIDANCE OF
Ms. PARUL TRIPATHI

NATIONAL MODEL
SENIOR SECONDARY SCHOOL
JANUARY
2025

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 CERTIFICATE

This is to certify that S.I. Vishali of class XII - PACIFIC

has prepared the report on the project titled “Gene
Manipulation”. The report is the result of her efforts and
endeavors. The report is found worthy of acceptance as the
final project report for the subject Biology.

Countersigned Signature of the Staff

Head of the Institution

Seal External Examiner

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 DECLARATION

I hereby declare that the project work entitled “Gene

Manipulation” submitted to the Department of Biology of
National Model Senior Secondary School, Peelamedu,
Coimbatore is a bonafide work of mine.

SIGNATURE OF THE STUDENT

NAME OF THE STUDENT

CLASS AND SECTION

REGISTER NUMBER

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 ACKNOWLEDGEMENT
I would like to express my sincere thanks and gratitude to my
teacher Ms. Parul Tripathi as well as our beloved Principal
Dr. D. Geetha who gave me the golden opportunity, support
and guidance to do this wonderful project on the topic “Gene
Manipulation”. Secondly, I would also like to thank my
parents and friends who helped me a lot in finishing this
project within the limited time.

Thanks again to all who helped me.

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 ABSTRACT

Genetic Manipulation is the direct manipulation of an organism’s genome

using biotechnology. It is a set of technologies used to change the genetic
makeup of cells, including the transfer of genes within and across species
boundaries to produce improved organisms. New DNA may be inserted in the
host genome by first isolating and copying the genetic material of interest using
molecular cloning methods to generate a DNA sequence, or by synthesising the
DNA and then inserting this construct into the host organisms.

GM techniques have been applied in numerous fields including research,

agriculture, industrial biotechnology and medicine. Enzymes used in laundry
detergent and medicine such as insulin and human growth hormone are now
manufactured in GM cells. GM animals such as mice or zebra fish are being
used for research purposes, and genetically modified crops have been
commercialised. Currently, biotechnology is looked upon as one of the most
promising branches of science.

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 TABLE OF CONTENTS

Chapter No. Title Page No.

Abstract v

1 Introduction 1

2 Genetically Modified Organisms 2

★ Genetically Modified Microbes
★ Genetically Modified Crops
★ Transgenic Animals
3 Genetic Engineering in Medicine 8
★ Artificial Blood
★ Cloned Pigs
★ Genetically Engineered FSH

4 Bio-fuel – As an Alternative 13

5 Bio-weapon – A Challenge 14

6 Conclusion 15

7 Bibliography 16

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 CHAPTER 1

INTRODUCTION
Genetic engineering is a process that alters the genetic structure of an
organism by either removing or introducing DNA, or modifying existing genetic
material in situ. Unlike traditional animal and plant breeding, which involves
doing multiple crosses and then selecting for the organism with the desired
phenotype, genetic engineering takes the gene directly from one organism and
delivers it to the other. This is much faster, can be used to insert any genes from
any organism and prevents other undesirable genes from also being added.

An organism that is generated through genetic engineering is considered

to be a genetically modified organism (GMO). The first GMOs were bacteria
generated in 1973 and GM mice in 1974. Insulin-producing bacteria were
commercialised in 1982 and genetically modified food has been sold since
1994. Glow fish, the first GMO designed as a pet, was first sold in the United
States in December 2003.

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 CHAPTER 2

GENETICALLY MODIFIED ORGANISMS

A genetically modified organism (GMO) or genetically engineered organism

is an organism whose genetic material has been altered using genetic
engineering techniques. Plants, animals or microorganisms that have changed
through genetic engineering are termed genetically modified organisms or
GMOs. Bacteria were the first organisms to be genetically modified. Plasmid
DNA containing new genes can be inserted into the bacterial cell and the
bacteria will then express those genes.

These genes can code for medicines or enzymes that process food and other
substrates. Plants have been modified for insect protection, herbicide
resistance,virus resistance, enhanced nutrition, tolerance to environmental
pressures and the production of edible vaccines. Most commercialised GMO's
are insect resistant and/or herbicide tolerant crop plants. Genetically modified
animals have been used for research, model animals and the production of
agricultural or pharmaceutical products. They include animals with genes
knocked out, increased susceptibility to disease, hormones for extra growth and
the ability to express proteins in their milk.

GMOs are used in biological and medical research, production of

pharmaceutical drugs, experimental medicine (e.g. gene therapy), and
agriculture(e.g. golden rice). For example, a gene from a jellyfish, encoding a
fluorescent protein called GFP,can be physically linked and thus co-expressed
with mammalian genes to identify the location of the protein encoded by the
GFP-tagged gene in the mammalian cell.

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➢ Genetically Modified Microbes:

Bacteria were the first organisms to be modified in the laboratory, due to their
simple genetics. These organisms are now used for several purposes, and are
particularly important in producing large amounts of pure human proteins for
use in medicine. Genetically, they are used to produce the protein insulin to treat
diabetes. Similar bacteria have been used to produce clotting factors to treat
haemophilia and human growth hormone to treat various forms of dwarfism.
Bacteria synthesise products such as:

● Insulin
● Hepatitis B vaccine
● Tissue plasminogen activator
● Human growth hormone
● Ice-minus bacteria
● Interferon

Gene therapy uses genetically modified viruses to deliver genes that can cure
disease into human cells. It has been used to treat genetic disorders such as
severe combined immunodeficiency. In 2004, researchers reported that a
genetically-modified virus that exploits the selfish behaviour of cancer cells
might offer an alternative way of killing tumours.

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 ➢ Genetically Modified Crops:

Transgenic plants have been engineered to possess several desirable traits, such
as resistance to pests, herbicides, or harsh environmental conditions, improved
product shelf life, and increased nutritional value. Since the first commercial
cultivation of genetically modified plants in 1996, they have been modified to
be tolerant to the herbicides glufosinate and glyphosate, to be resistant to virus
damage as in Ringspot virus-resistant GM papaya, grown in Hawaii, and to
produce the Bt toxin, an insecticide that is non-toxic to mammals. Most GM
crops grown today have been modified with "input traits", which provide
benefits mainly to farmers.

● Golden Rice :

It is a transgenic variety of rice, with genes for the synthesis of b-carotene

taken from the temperate garden favourite Narcissus pseudonarcissus (daffodil)
and inserted into the genome of a temperate strain of rice, using Agrobacterium
tumefaciens as the vector, to effect the transfer. The gene construct also contains
some genes for enzymes of the biosynthetic pathway of b-carotene, from
another bacterium Erwinia uredovora.

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 ➢ Transgenic Animals:

A transgenic animal is one that carries a foreign gene that has been
deliberately inserted into its genome. The foreign gene is constructed using
recombinant DNA methodology. Transgenic animals are used as experimental
models to perform phenotypic and for testing in biomedical research.
Genetically Modified (Genetically Engineered) animals are becoming more vital
to the discovery and development of cures and treatments for many serious
diseases. By altering the DNA or transferring DNA to an animal, we can
develop certain proteins that may be used in medical treatment. Stable
expressions of human proteins have been developed in many animals, including
sheep, pigs, and rats.

Some examples are: Human-alpha-1-antitrypsin, which has been developed

in sheep and is used in treating humans with this deficiency and transgenic pigs
with human-histo-compatibility have been studied in the hopes that the organs
will be suitable for transplant with less chances of rejection. Many medicines,
including insulin and many immunizations are developed in transgenic animals.
In March 2011, the bioactive recombinant Human Lysozyme was expressed in
the milk of cloned transgenic cattle. This field is growing rapidly and new
farming uses are being discovered and developed.

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 ● The Glow fish:

In 1999, Dr. Zhiyuan Gong and his colleagues at the National University of
Singapore were working with a gene called green fluorescent protein (GFP),
originally extracted from a jellyfish, that naturally produces bright green
fluorescence. They inserted the gene into a zebrafish embryo, allowing it to
integrate into the zebra fish’s genome, which caused the fish to be brightly
fluorescent under both natural white light and ultraviolet light. Their goal was to
develop a fish that could detect pollution by selectively fluorescing in the
presence of environmental toxins.The development of the constantly fluorescing
fish was the first step in this process.Shortly thereafter, his team developed a
line of red fluorescent zebrafish by adding a gene from a sea coral, and orange-
yellow fluorescent zebrafish, by adding a variant of the jellyfish gene. Later, a
team of researchers at the National University of Taiwan,succeeded in creating a
medaka (rice fish) with a fluorescent green colour, which like the zebrafish is a
model organism used in biology.

● Malaria Resistant Mosquitoes:

 Scientists have been exploring the use of genetically modified mosquitoes to
prevent the spread of malaria and other diseases. One challenge is ensuring that
the modified mosquitoes mate with wild mosquitoes and transfer the protective
trait to their offspring. A team led by Dr. George Dimopoulos at Johns Hopkins
University genetically modified Anopheles mosquitoes to boost immune activity
in their midguts. This immune boost successfully suppressed malaria-causing
Plasmodium parasites as well as bacteria.

The scientists showed that the immune boost caused changes in the microbial
community, or microbiota, of both the mosquito midgut and reproductive
organs. This, in turn, altered mating preferences among the mosquitoes.The
researchers caged equal numbers of wild and genetically modified mosquitoes
and then monitored their breeding. Genetically modified males preferred wild
females, and wild males preferred modified females. These preferences helped
to quickly spread the protective trait through the mosquito population.

CHAPTER 3
 GENETIC ENGINEERING IN MEDICINE

Genetic engineering is becoming a major force in conventional medicine. It

Has got numerous applications in medicine ranging from vaccines to transgenic
organ transplants.

➢ Artificial Blood:

The production of artificial blood can be done in a variety of ways. For

haemoglobin-based products, this involves isolation or synthesization of
haemoglobin, molecular modification then reconstitution in an artificial blood
formula.

Methods for Production:

● To obtain haemoglobin, a strain of E. coli bacteria that has the ability to

produce human haemoglobin is used. Over the course of about three days, the
protein is harvested and the bacteria are destroyed. To start the fermentation
process, a sample of the pure bacteria culture is transferred to a test tube that
contains all the nutrients necessary for growth. This initial inoculation causes
the bacteria to multiply. When the population is great enough, they are
transferred to a seed tank.
● A seed tank is a large stainless steel kettle that provides an ideal
environment for growing bacteria. It is filled with warm water, food, and an
ammonia source which are all required for the production of haemoglobin.
Other growth factors such as vitamins, amino acids, and minor nutrients are also
added. The bacterial solution inside the seed tank is constantly bathed with
compressed air and mixed to keep it moving. When enough time has passed, the
contents of the seed tank is pumped to the fermentation tank.
● 8
● The fermentation tank is a larger version of the seed tank. It is also filled
with a growth media needed for the bacteria to grow and produce haemoglobin.
Since pH control is vital for optimal growth, ammonia water is added to the tank
as necessary. When enough haemoglobin has been produced, the tank is emptied
so isolation can begin.
● Isolation begins with a centrifugal separator that isolates much of the
haemoglobin. It can be further segregated and purified using fractional
distillation. This standard column separation method is based on the principle of
boiling a liquid to separate one or more components and utilises vertical
structures called fractionating columns. From this column, the haemoglobin is
transferred to a final processing tank.
● Here, it is mixed with water and other electrolytes to produce the artificial
blood. The artificial blood can then be pasteurised and put into an appropriate
packaging. The quality of compounds is checked regularly during the entire
process.

➢ Cloned Pigs:
 Two competing teams have cloned pigs that have been genetically modified to
produce organs more suitable for transplantation into humans. Pig organs are
well suited for transplantation; they are approximately the same size as human
organs and have similar plumbing, which makes reconnecting blood vessels
much easier. Also, the size of pig litters tends to be large and pigs reproduce
quickly, raising the prospect of a large supply of "spare" organs.

A problem with using pig organs, however, is that they are coated with sugar
molecules that trigger acute rejection in people. Human antibodies attach
themselves to these sugar molecules and quickly destroy the newly transplanted
pig organ. To circumvent the rejection, scientists are working to produce pigs
that lack the sugar-producing gene.

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➢ Genetically Engineered FSH:

 FSH is produced by the pituitary gland and directly stimulates the ovaries to
recruit and support ovarian follicles, each containing one egg. The
hypothalamus adjusts the production of FSH depending upon the levels of other
hormones such as oestrogen. FSH is used in stimulated IUI and assisted
reproductive technology cycles (IVF) because it causes the development of
numerous follicles. More follicles are needed in ART cycles because some do
not fertilise or do not continue to develop.

FSH products are divided into two categories:

1) Those extracted from the urine of postmenopausal women:

Repronex, Menopur and Bravelle

2) Those made using genetic recombinant technology:

Gonal-F and Follistim.

● Urinary FSH Products (Repronex, Bravelle, Menopur):

FSH is excreted in the urine and levels increase after menopause because of the
lack of good quality eggs. These products are made by extracting and purifying
the urine from postmenopausal women. Repronex and Menopur contain equal
amounts of FSH and luteinizing hormone (LH) whereas Bravelle is further
purified to remove all but trace amounts of LH.

These products are reconstituted from a powder and given with a syringe either
intramuscularly or subcutaneously. The manufacturers provide recommended
doses, however, doses vary based on each patient’s response and treatment
history.

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 ● Recombinant Products (Gonal-F, Follistim):

Gonal-F and Follistim contain 75 IU of FSH in each vial and are made by
cloning the gene which codes for human FSH. The gene is inserted into
mammalian cell cultures which then produce FSH. These drugs are given
subcutaneously using a specially designed pen.

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CHAPTER 4
 BIOFUEL- AS AN ALTERNATIVE

Algae Fuel – It might be an alternative to fossil fuel and uses algae as its
source of natural deposits. Several companies and government agencies are
funding efforts to reduce capital and operating costs and make algae fuel
production commercially viable. Harvested algae, like fossil fuel, release CO2
when burnt but unlike fossil fuel the CO2 is taken out of the atmosphere by the
growing algae. High oil prices, competing demands between foods and other
biofuel sources, and the world food crisis, have ignited interest in algal
culture(farming algae) for making biodiesel, bioethanol, biogasoline,
biomethanol, biobutanol and other biofuels, using land that is not suitable for
agriculture.

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CHAPTER 5
 BIOWEAPON- A CHALLENGE

Rapid developments in biotechnology, genetics and genomics are

undoubtedly creating a variety of environmental, ethical, political and social
challenges for advanced societies. But they also have severe implications for
international peace and security because they open up tremendous avenues for
the creation of new biological weapons.
The history of biological warfare is nearly as old as the history of warfare
itself. In ancient times, warring parties poisoned wells or used arrowheads with
natural toxins. By using genetic engineering, biological researchers have already
developed new weapons that are much more effective than their natural
counterparts.

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CHAPTER 6
 CONCLUSION

Promising aspects of Genetic Engineering that can bring about tremendous

changes in human life is well known. However the manipulation of living
organisms by the human race cannot go on any further without regulation. Some
ethical standards are required to evaluate the morality of all human activities
that might help or harm living organisms. Going beyond the morality of such
issues the biological significance of such things is also important. Genetic
Modification of organisms can have unpredictable results when such organisms
are introduced into the ecosystem.

Every new technology aims to improve man’s life. It is for man to

make judicious use of its applications!

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BIBLIOGRAPHY
1. www.wikipedia.com
2. www.encyclopedia.com
3. Britannica Encyclopedia
4. Tell Me Why Magazine
5. Academia.edu
6. Study.com

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