BIOLOGY INVESTIGATORY PROJECT
CLASS 12 (2023-2024)
G e
netic Engineering
Applications
Dheeptha Krishnamurthy
Genetic Engineering Applications
XII NEET
The century that we left behind has witnessed giant
strides in almost all spheres of human life. Currently,
biotechnology is looked upon as one of the most promising
branches of science. And it is Genetic Engineering that
makes most biotechnological applications possible.
Genetic engineering is the direct human manipulation of
an organism's genome using modern DNA technology. It
involves the introduction of foreign DNA or synthetic genes
into the organism of interest. The introduction of new DNA
does not require the use of classical genetic methods;
however traditional breeding methods are typically used
for the propagation of recombinant organisms.
Humans have altered the genomes of species for
thousands of years through artificial selection and more
recently mutagenesis. Genetic engineering as the direct
manipulation of DNA by humans outside breeding and
mutations has only existed since the 1970s. Humans have
altered the genomes of species for thousands of years
through artificial selection and more recently mutagenesis.
Genetic engineering as the direct manipulation of DNA by
humans outside breeding and mutations has only existed
since the 1970s. The most common form of genetic
engineering involves the insertion of new genetic material
at an unspecified location in the host genome. This is
accomplished by isolating and copying the genetic
material of interest using molecular cloning methods to
generate a DNA sequence containing the required genetic
elements for expression, and then inserting this construct
into the host organism. Other forms of genetic engineering
include gene targeting and knocking out specific genes via
the most common form of genetic engineering involves
the insertion of new genetic material at an unspecified
location in the host genome. This is accomplished by
isolating and copying the genetic material of interest using
molecular cloning methods to generate a DNA sequence
containing the required genetic elements for expression,
and then inserting this construct into the host organism.
Genetic engineering alters the genetic makeup of an
organism using techniques that introduce heritable
material prepared outside the organism either directly into
the host or into a cell that is then fused or hybridized with
the host. This involves using recombinant nucleic acid
(DNA or RNA) techniques to form new combinations of
heritable genetic material followed by the incorporation of
that material either indirectly through a vector system or
directly through micro-injection, macro-injection and
micro- encapsulation techniques.
Genetic engineering techniques have been applied
in numerous fields including research, agriculture,
industry, and medicine. This project seeks to focus on
some most promising areas of biotechnological
applications...
GENETICALLY MODIFIED ORGANISMS
A genetically modified organism (GMO) or
genetically engineered organism (GEO) is an organism
whose genetic material has been altered using genetic
engineering techniques. These techniques, generally
known as recombinant DNA technology, use DNA
molecules from different sources, which are combined into
one molecule to create a new set of genes. This DNA is
then transferred into an organism, giving it modified or
novel genes. Transgenic organisms, a subset of GMOs, are
organisms which have inserted DNA that originated in a
different species.
GMOs are used in biological and medical research,
production of pharmaceutical drugs, experimental
medicine (e.g. gene therapy), and agriculture (e.g. golden
rice). The term "genetically modified organism" does not
always imply, but can include, targeted insertions of genes
from one species into another. 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. Such
methods are useful tools for biologists in many areas of
research, including those who study the mechanisms of
human and other diseases or fundamental biological
processes in eukaryotic or prokaryotic cells.
a) 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 are used to
produce the protein insulin to treat diabetes. Similar
bacteria have been used to produce clotting factors to
treat hemophilia and human growth hormone to treat
various forms of dwarfism. Bacteria synthesize products
such as;
Insulin
Hepatitis B vaccine
Tissue plasminogen activator
Human growth hormone
Ice-minus bacteria
Interferons
In materials science, a genetically modified virus has been
used to construct more environmentally friendly lithium-
ion battery.
Gene therapy uses genetically modified viruses to deliver
genes that can cure disease into human cells. Although
gene therapy is still relatively new, it has had some
successes. 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.
Genetically modified virus and bacteria
b) 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 Ring spot 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 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.
Genetically modified foods (GM foods or GMO foods)
are foods derived from genetically modified organisms
(GMOs). Genetically modified organisms have had specific
changes introduced into their DNA by genetic engineering
techniques. These techniques are much more precise than
mutagenesis (mutation breeding) where an organism is
exposed to radiation or chemicals to create a non-specific
but stable change. Other techniques by which humans
modify food organisms include selective breeding; plant
breeding, and animal breeding, and soma-clonal variation.
GM foods were first put on the market in 1996.
Typically, genetically modified foods are transgenic plant
products: soybean, corn, canola, and cotton seed oil.
Animal products have also been developed, although as of
July 2010 none are currently on the market! In 2006 a pig
was controversially engineered to produce omega-3 fatty
acids through the expression of a roundworm gene.
Researchers have also developed a genetically-modified
breed of pigs that are able to absorb plant phosphorus
more efficiently, and as a consequence the phosphorus
content of their manure is reduced by as much as 60%.La!
Critics have objected to GM foods on several grounds,
including safety issues ecological concerns, and
economic concerns raised by the fact these organisms
are subject to intellectual property law.
c) 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. In addition to the gene itself, the DNA
usually includes other sequences to enable it to be
incorporated into the DNA of the host and to be expressed
correctly by the cells of the host.
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. Transgenic livestock have been used
as bioreactors since the 1990s. 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 pharming uses are being discovered and
developed. The extent that transgenic animals will be
useful in the medical field as well as other fields is very
promising based on results thus far.
The Glow Fish, a fluorescent red zebra fish sold as a
novel pet, has become the first transgenic animal sold to
U.S. consumers. 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 froma jellyfish, that naturally
produced bright green fluorescence. They inserted the
gene into a zebra fish embryo, allowing it to integrate into
the zebrafish’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 zebra fish by
adding a gene from a sea coral, and orange-yellow
fluorescent zebra fish, by adding a variant of the jellyfish
gene.
Later,
a team of researchers at the National University of Taiwan,
headed by Professor Huai-Jen Tsai succeeded in creating a
medaka (rice fish) with a fluorescent green color, which
like the zebrafish is a model organism used in biology.
Fruit flies
In biological research, transgenic fruit flies (Drosophila
melanogaster) are model organisms used to study the
effects of genetic changes on development.! Fruit flies are
often preferred over other animals due to their short life
cycle, low maintenance requirements, and relatively simple
genome compared to many vertebrates.
Mosquitoes
In 2010, scientists created “malaria-resistant mosquitoes”
in the laboratory. The World Health Organization estimated
that Malaria killed almost one million people in 2008.
Genetically modified male mosquitoes containing a lethal
gene have been developed in order to combat the spread
of Dengue fever. Aedes aegypti mosquitoes, the single
most important carrier of dengue fever, were reduced by
80% in a 2010 trial of these GM mosquitoes in the Cayman
Islands. Around 50 - 100 million people are affected by
Dengue fever every year and 40,000 people die from it.
Mammals
Genetically modified mammals are an important category
of genetically modified organisms. Transgenic mice are
often used to study cellular and tissue-specific responses
to disease.
In 1999, scientists at the University of Guelph in Ontario,
Canada created the genetically engineered Enviropig. The
Enviropig excretes from 30 to 70.7% less phosphorus in
manure depending upon the age and dietlIn February
2010, Environment Canada determined that Enviropigs are
in compliance with the Canadian Environmental Protection
Act and can be produced outside of the research context in
controlled facilities where they are segregated from other
animals.
In 2009, scientists in Japan announced that they had
successfully transferred a gene into a primate species
(marmosets) and produced a stable line of breeding
transgenic primates for the first time. Their first research
target for these marmosets was Parkinson's disease, but
they were also considering Amyotrophic lateral sclerosis
and Huntington's discase.
In 2011, scientists in China released news that they have
introduced human genes into 300 dairy cows to produce
milk with the same properties as human breast milk. Aside
fram milk production, the researchers claim these
transgenic cows to be identical to regular cows,
Cnidarians
Cnidarians such as Hydra and the sea anemone
Nematostella vectensis have become attractive model
organisms to study the cvolution of immunity and certain
developmental processes. An important technical
breakthrough was the development of procedures for
generation of stably transgenic hydras and sea anemones
by embryo microinjection.
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.
a) The Artificial Blood
The artificial blood is a genetically engineered form
of hemoglobin, the complicated protein that enclosed in
red blood cells--carries oxygen from the lungs to tissues
throughout the body. Many companies have been
searching for an artificial blood because of the annual
worldwide shortage of about 100 million units of blood and
the military's need for blood replacements that can be
stored in field conditions without refrigeration. An artificial
blood also would virtually eliminate the risk of contracting
AIDS, hepatitis and other viral diseases through
transfusions.
Development of emulsion technologies resulted in
the production of compounds which utilized smaller chain
perfluorocarbon molecules to more effectively emulsify the
perfluorocarbons, allowing higher concentrations of active
agent in the emulsion and thus higher oxygen carrying
capabilities. The improved stability of the newer emulsions
are vastly superior to the first generations of
perfluorocarbons; current emulsions can be stored at 4°C
for extended periods of time (months) without appreciable
degradation of activity.
b) Cloned Pigs
Modified for Use in Human Transplants
Two competing teams have cloned pigs that have heen
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.
c) 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 estrogen. 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 fertilize or do not continue to develop. There
are several FSH products available in the United States
including Follistim, Gonal-F, Bravelle, and Repronex.
Follistim and Gonal-F are produced by genetic engineering
using mammalian culture cells. These products are
identical to the body's FSH. Bravelle and Repronex are
derived from the urine of postmenopausal women.
BIOFUEL- AN ALTERNATIVE
Algae fuel 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 alga culture(farming algae) for making
biodiesel, bioethanol, biogasoline, biomethanol, biobutanol
and other biofuels, using land that is not suitable for
agriculture
Biodiesel
Currently most research into efficient algal-oil production
is being done in the private sector, but predictions from
small scale production experiments bear out that using
algae to produce biodiesel may be the only viable method
by which to produce enough automotive fuel to replace
current world diesel usage.= If algae-derived biodiesel
were to replace the annual global production of 1.lbn tons
of conventional diesel, a land mass of 57.3 million
hectares would be required. This compares highly
favourable to other biofuelsila Microalgae have much
faster growth rates than terrestrial crops. The per unit area
yield of oil from algae is estimated to be from between
5,000 to 20,000 US gallons per acre per year (4,700 to
18,000 m3/1cm2.a).11LI This is 7 to 30 times greater than
the next best crop, Chinese tallow (700 US gal/acre•a or
650 m3/Itin•a).
Studies show that some species of algae can produce up
to 60% of their dry weight in the form of oiL Because the
cells grow in aqueous suspension, where they have more
efficient access to water, CO2 and dissolved nutrients,
microalgae are capable of producing large amounts of
biomass and usable oil in either high rate algal ponds or
photo bioreactors. This oil can then be turned Into
biodiesel which could be sold for use In automobiles.
Regional production of microalgae and processing into
biofuels will provide economic benefits to rural
communities.
The National Algae Biofuel Technology Roadmap
emphasizes the role of genetic engineering to find, adjust
and deploy optimized algae species for biofuel
development.
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 genetically engineered 'superbug'—highly
lethal and resistant to environmental influence or any
medical treatment—is only a small part of this story. Much
more alarming, from an arms-control perspective, are the
possibilities of developing completely novel weapons on
the basis of knowledge provided by biomedical research—
developments that are already taking place. Such
weapons, designed for new types of conflicts and warfare
scenarios, secret operations or sabotage activities, are not
mere science fiction, but are increasingly becoming a
reality that we have to face.
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. Countless examples from
the daily work of molecular biologists could be presented
here, not least the introduction of antibiotic resistance into
bacterial pathogens, which today is routine work in almost
any microbiology laboratory. Indeed, many research
projects in basic science show—sometimes unwillingly and
unwittingly—how to overcome current scientific and
technological limits in the military use of pathogenic
agents. Furthermore, genetic engineering is not merely a
theoretical possibility for future biowarfare: it has already
been applied in past weapons programmes, particularly in
the former Soviet Union. One example is the USSR's
'invisible anthrax', resulting from the introduction of an
alien gene into Bacillus anthracis that altered its
immunological properties (Pnmerantsev eta!,. 1997).
Existing vaccines proved to be ineffective against this new
genetically engineered strain.
The global norm against biological weapons, laid down in
the 1925 Geneva Convention and the 1972 Biological and
Toxin Weapons Convention, clearly contributed to the fact
that few countries have been engaged in research into
offensive biowarfare during recent decades. This moral
barrier seems to be lower for 'non-lethal' weapons that are
targeted against materials or drug-producing plants.
Indeed, today's technical possibilities are creating a new
interest in this area that might be leading to a new
biological arms race.
New technological possibilities met new military concepts
in the USA and led to a renewed interest in weapons that,
until recently, had been banned and rejected.
In 1998, it became public that the US Naval Research
Laboratory in Washington DC was developing genetically
engineered fungi with offensive biowarfare potential. They
isolated natural microorganisms that degrade a variety of
materials, such as plastics, rubber and metals, and used
genetic engineering to make them more powerful and
focused—one of these genetically engineered microbes
can destroy military paints in 72 hours.
Microbes that are composed of synthetic or artificial
components, such as DNA or RNA or cockA or amino acids
can be tailored for use as biological warfare agents. As
examples, if DNA used x1211A were to be coupled with
synthetic codons, with synthetic anti-codons, etc. then
such microbes used as biological warfare agents would be
synthetic biological warfare agents.
CONCLUSION
We have discussed the promising aspects of Genetic
Engineering that can bring about tremendous changes in
human life. 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 the Judicious
use of its applications...
