JESUS NEVER FAILS

St. PAUL PUBLIC SCHOOL – CBSE

SENIOR SECONDARY
PANIKANKUPPAM, PANRUTI.

2023 – 2024
PROJECT
NAME :

STD :

ROLL NO :

SUBJECT :

TOPIC :

UNDER THE GUIDANCE OF

SIGNATURE OF THE TEACHER SIGNATURE OF THE PRINCIPAL

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 St. PAUL PUBLIC SCHOOL – CBSE
SENIOR SECONDARY

CERTIFICATE

This is to certify that ________________________________ , a student

of class XII has successfully completed the research on the

below mentioned project ____________________________________

________________________________________________________

Under the guidance of

______________________________________

during the year 2023-2024 in partial fulfilment of practical examination

conducted by AISSCE, NEW DELHI.

Signature of the Internal Examiner Signature of the External Examiner

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S.NO TABLE OF CONTENTS PAGE NO

1. ABSTRACT

2. GENETIC ENGINEERING

3. GENETIC ENGINEERING HAS PERMITTED

4. RECOMBINANT DNA TECHNIQUE

5. GENETICALLY ENGINEERED INSULIN

6. GENE THERAPY

7. CONCLUSION

8. BIBLIOGRAPHY

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CRISPR

CRISPR technology or CRISPR-Cas9 utilizes a protein called Cas9,

which acts like a pair of molecular scissors and can cut DNA.
CRISPRs are specialized stretches of DNA and are used in medical
biotechnology as a tool to edit genomes. This allows scientists to
alter DNA and modify gene functions, often called genetic
engineering. There are many applications, like correcting genetic
defects, treating diseases, preventing the spread of diseases,
improving crops, and more. But the science of altering genomes has
many ethical concerns surrounding it. From the ability to mutate
genes and the unknowns surrounding gene mutation, CRISPR is a
controversial area of biomedicalscience. Some new studies even
show that perhaps CRISPR technology can create tumors and cancer
with DNA deletions that aren’t controlled or precise. Of course,
pharmaceutical companies and other scientific organizations that
develop and utilize CRISPR technology are trying to downplay the
concerns and issues, so the reality of the benefits and
damage of the technology is somewhat unknown.

STEM CELL RESEARCH

Biotechnology plays a big part in supporting stem cell research,

which supports the exploration of growing stem cells in a lab setting
or in vitro. This could help in situations where patients may be
suffering from a disease or disorder where implanting stem cells
could help restore their vitality and give them a new lease on life.
How does it work? Because stem cells can repeatedly divide and
transform into other types of body cells, biotechnologists can learn
how to work with their unique profiles to encourage growth of
specific types of cells. Though research is ongoing, it’s reported that
the results show hope for the future of this unique medical approach.
Genetically Engineered Insulin (Humulin)
Insulin is a peptide hormone produced
by beta cells in the pancreas of various
organisms including human beings. It
regulates
the metabolism of carbohydrates an
d fats by promoting the absorptionof
glucose from the blood to skeletal
muscles and fat tissue and by causing
fat to be stored rather than used for energy. Insulin also inhibits the
production of glucose by the liver.

Except in the presence of the metabolic disorder diabetes mellitus

and metabolic syndrome, insulin is provided within the body ina constant
proportion to remove excess glucose from the blood, which otherwise
would be toxic. When blood glucose levels fall below a certain level, the
body begins to use stored glucose as an energy source through
glycogenolysis, which breaks down the glycogen stored in the liver and
muscles into glucose, which can then be utilized as an energysource. As
a central metabolic control mechanism, its status is also usedas a control
signal to other body systems (such as amino acid uptake bybody cells).
In addition, it has several other anabolic effects throughout the body.
When control of insulin levels fails, diabetes mellitus can result.

Structure:

Insulin is composed of two

different types of peptide
chains. Chain A has 21 amino
acids and Chain B has 30 amino
acids. Both chains contain alpha
helices but no beta strands.
There are 3 conserved disulfide
bridges which help keep the
two chains together. Insulin can
also form dimers in solution due
to the hydrogen bonding between the B chains. The dimers can further
interact to form hexamers due to interaction between hydrophobic
surfaces. This scene highlights the hydrophobic and polar parts of an insulin
monomer at a pH of 7.

A number of insulin variants have been made to favor either the

monomeric or hexameric form. Deletion of the five C terminal residues
of the B chain creates a monomer only form. This portion of the B chain
is involved in hydrogen bonds between the B chain of one monomer and
the A and B chain of another monomer.

Need of Genetically Engineered Insulin:

The original form of the wonder cure for diabetes, these were once the
only type of insulin available, but are now rarely used. Animal insulin was
originally made
from ground-up
animal pancreas
tissue, and then later
was extracted from
healthy animals
(slaughtered pigs &
cows). The
metabolism of cows and pigs was close enough to human metabolism that
their animal insulin also worked well in human bodies. Beef insulin has 3
differences from human; pork insulin has 1 difference from human. The
use of a mixture of beef and pork insulin was also possible.It has been
shown that human insulin is less immunogenic than animal insulin. Porcine
insulin is most similar to human insulin. The primary amino acid sequences
of bovine and porcine insulin differ from that of human insulin by three
and one amino acid, respectively. This greater dissimilarity between
human and bovine insulin has been postulated tobe the explanation for
the greater antigenicity of bovine insulin as compared with porcine insulin

One of the problems with animal insulin was antibody issues. The body
identifies them and tries to reject them. Pork insulin differs by 1 amino
acid and beef insulin by 3 amino acids, so the body's immune system can
sometimes recognize them as foreign. Immunological complications of
insulin therapy have been evident since animal insulin became available
for the treatment of diabetes mellitus in 1922. In insulin-allergic patients
treated with conventional insulin preparations,
the insulin-specific IgE values are often 10- to 20-fold higher than in
patients without allergy. It has been shown that human insulin is less
immunogenic than animal insulin. Porcine
insulin is most similar to human insulin. Cross-
reactivity between human insulin and insulin
of animal origin has been reported. A major
problem is the cross-reactivity that occurs
between anti-insulin antibodies and thevarious
animal and human insulin preparations in
patients presenting with allergy to animal
insulin.

The usage of animal insulin has so greatly declined in modern times

that they have largely been withdrawn from the market. Newly
diagnosed diabetics are typically given synthesized or Genetically
Engineered human insulin.

What is “Proinsulin”?

Proinsulin is the prohormone precursor to insulin made in the beta cells

of the islets of Langerhans, specialized regions of the pancreas.
Proinsulin is synthesized on
membrane associated
ribosomes found on the rough
endoplasmic reticulum, where it
is folded and its disulfidebonds
are oxidized. It is then
transported to the Golgi
apparatus where it is packaged
into secretory vesicles, and
where it is processed by a series
of proteases to form
mature insulin. Mature insulin has 35 fewer amino acids; 4 are removed
altogether, and the remaining 31 form the C-peptide. The C-peptide is
abstracted from the center of the proinsulin sequence; the two other ends
(the B chain and A chain) remain connected by disulfide bonds.

When insulin was originally purified from bovine or porcine pancreata,

all the proinsulin was not fully removed. [3][4] When some people used these
insulins, the proinsulin may have caused the body to react with a
rash, to resist the insulin, or even to make dents or lumps in the skin at
the place where the insulin was injected. This can be described as an
iatrogenic injury due to slight differences between the proinsulin of
different species. Since the late 1970s, when highly purified
porcine insulin was introduced, and the level of insulin purity reached 99%,
this ceased to be a significant clinical issue. The main challenge for
production of insulin using rDNA techniques was getting insulin
assembled into mature form.

Humulin:

Humulin was the first medication produced using modern genetic

engineering techniques in which actual human DNA is inserted into a
host cell (E. coli in this case). Biosynthetic "human" insulin is now
manufactured for widespread clinical use using genetic engineering
techniques using recombinant DNA technology, which themanufacturers
claim reduces the presence of many impurities, although there is no
clinical evidence to substantiate this claim. EliLilly marketed the
first artificial insulin, Humulin, in 1982.

Humulin production method is as follows:

1. DNA coding for A and B polypeptide chains of insulin are chemically

synthesised a in the lab. Sixty three nucleotides are sequenced to
produce A chain of insulin and ninety nucleotide long DNA designed
to produce B chain of insulin, plus terminator codonis added at the
end of each chain sequence. Anti-codon for methionine is added at
the beginning of the sequence to distinguish humulin from the other
bacterial proteins.

2. Chemically synthesized A and B chain DNA sequence are inserted

into one of the marker gene which are present in the plasmid
vector. Genes are inserted into the plasmid with the help of
enzymes known as endonuclease and ligase.

3. The vector plasmids with the insulin gene are then introduced into
the E. coli bacterial cell. These cells are then allowed to replicate
by mitosis, along with the bacterial cell recombinant plasmid also
gets replicated producing the human insulin.

4. A and B polypeptide chains of insulin are then extracted and purified

from the fomenters in the lab. High-Performance Liquid
 Chromatography (HPLC) is used to get 100% pure humulin from
the mixture of proteins.

5. The A and B polypeptide chains of insulin are mixed together and

connected with each other by disulphide bond, forming the Humulin
or synthetic human insulin.

Advantages & Disadvantages of Humulin:

Humulin is the one and only human protein produced in the bacteria
with identical chemical structure to that of the natural human insulin.
Administration of humulin reduces the possibility of antibody production
and inflammatory response
in diabetic patients. Major
difficulty is the extraction of
humulin from a mixture of
host proteins present in the
fermentation broth.

Now days to overcome this

extraction problem synthetic
human insulin are produced
in the yeast cell instead of E. coli using the same procedure. As yeast is
Eukaryotes they secrete the whole humulin molecule with perfect three
dimensional structures, reducing the need for complex and time
consuming purification methods.

Now most of the diabetic patients are treated with synthetic human
insulin. Small group of patients claim that episodes of hyperglycaemic
complications have been increased after shifting from animal origin
insulin to humulin. No study till date shows the difference between the
frequency of hyperglycaemic complications in patient using humulin
(synthetic human insulin) and animal origin insulin.
Gene Therapy
Gene therapy is the therapeutic delivery of nucleic acid polymers into
a patient's cells as a drug to treat disease. Gene therapy is anexperimental
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 only
being tested for the treatment of diseases that have no other cures. It
should be noted that not all medical procedures that introduce alterations
to a patient's genetic makeup can be considered gene therapy. Bone
marrow transplantation, and organ transplants in general have been found
to introduce foreign DNA into patients. Gene therapy is defined by the
precision of the procedure and the intention of direct therapeutic effects.

Gene therapy was conceptualized in 1972, by authors who urgedcaution

before commencing human gene therapy studies.

The first attempt, albeit 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.

The first germ line gene therapy consisted of producing a genetically

engineered embryo in October 1996. The baby was born on July 21,
1997 and was produced by taking a donor's egg with healthy mitochondria,
removing its nuclear DNA and filling it with the nuclear DNA of the
biological mother - a procedure known as cytoplasmic transfer.

This procedure was referred to sensationally and somewhat inaccurately

in the media as a "three parent baby", though mtDNA is not the primary
human genome and has little effect on an organism's individual
characteristics beyond powering their cells.

Gene therapy is a way to fix a genetic problem at its source. Thepolymers

are either expressed as proteins, interfere with protein 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.

The first commercial gene therapy, Gendicine, was approved in China in

2003 for the treatment of certain cancers. In 2011 Neovasculgen was
registered in Russia as the first-in-class gene-therapy drug for treatment
of peripheral artery disease, including critical limb ischemia. In2012
Glybera, a treatment for a rare inherited disorder, became the first
treatment to be approved for clinical use in either Europe or the United
States after its endorsement by the European Commission.

ADA deficiency is one form of SCID (severe combined immunodeficiency),

a disorder that affects the immune system. ADAdeficiency is very rare, but
very dangerous, because a malfunctioning immune system leaves the body
open to infection from bacteria and viruses.
The disease is caused by a
mutation in a gene on
chromosome 20. ADA
deficiency is inherited in
an autosomal recessive
manner. The gene codes for
the enzyme adenosine
deaminase (ADA). Without
this enzyme, the body is
unable to break down a toxic
substance called
deoxyadenosine. The toxin
builds up and destroys
infection-fighting immune
cells called T and B
lymphocytes. Because ADA
deficiency affects the
immune system, people who have the disorder are more susceptible to
all kinds of infections, particularly those of the skin, respiratory system,
and gastrointestinal tract. They may also be shorter than normal. Sadly,
most babies who are born with the disorder die within a few months.

Treatments of ADA Deficiency includes:

• bone marrow transplant

• gene therapy

• ADA enzyme in PEG vehicle

On September 14, 1990, the first gene therapy to combat this disease
was performed by Dr. William French Anderson on a four-year-old girl,
Ashanti DeSilva, at the National Institutes of Health, Bethesda,
Maryland, U.S.A.
 Conclusion
Biotechnology is the new wonder of science. It is truly
multidisciplinary in nature and it encompasses several disciplines of
basic sciences and engineering. The Science disciplines from which
biotechnology draws heavily are microbiology, chemistry,
biochemistry, genetics, molecular biology, immunology, cell and
tissue culture and physiology. On theengineering side it leans heavily
on process chemical and biochemical engineering since large scale
cultivation of microorganisms and cells, their downstream processing
are based on them. It comes to us as a great blessing.

Biotechnology utilizes the technique called genetic engineering or

recombinant DNA technology where a microorganism is isolated;
its genetic material is cut, manipulated, sealed, again inserted in
an organism and allowed to grow in a suitable environment under
controlled conditions to get the desired product. It looks easy but
is a very tedious job and it takes years for a research to achieve its
goal.
Like every other thing, biotechnology too has some harmful
impacts:
1. Genetic engineering is a very vital part of biotechnology and the
cost of transferring genes from one species to another is very
expensive, which requires a huge amount of capital investment.
The cost of producing genetically- modified plants and animals
are sky- rocketing and the duration of return are also not
predictable.
2. Genetic engineering crosses boundaries of reproduction by crossing
genes of species that are completely unrelated; hencegiving rise to
hazardous results as well as also increasing the riskof harming
multiple species.
3. When genetic material from certain viruses is used in the
production of transgenic crops, there are chances that these virus
genes will combine with crop genes to produce more destructive
viruses. The consumption of such crops is hazardous to human
health and can cause several life- threatening ailments. It can also
result in cancer, often malignant as well.
4. Biotechnology also poses a number of environmental threats.
Genetically modifies crops often infect monarch butteries and other
insect species.
 The applications of biotechnology are so broad, and the advantages
so compelling, that virtually every industry is using this technology.
Developments are underway in areas as diverse as pharmaceuticals,
diagnostics, textiles, aquaculture, forestry, chemicals, household
products, environmental cleanup, food processing and forensics to name
a few. Biotechnology is enabling these industries to make new or better
products, often with greater speed, efficiency and flexibility.
Biotechnology must continue to be carefully regulated so that the
maximum benefits are received with the least risk.

Bibliography
http://en.wikipedia.org/biotechnology
http://en.wikipedia.org/insulin
http://www.genewatch.org/sub-568238
http://en.wikipedia.org/humulin
http://www.biotecharticles.com/Others-Article/Human-

Insulin-and-Recombinant-DNA-Technology-70.html
https://isaaa.org/resources/publications/pocketk/34/default.

asp
http://www.sciencedirect.com/
https://en.wikipedia.org/wiki/Gene_therapy
https://en.wikipedia.org/wiki/Adenosine_deaminase_deficie

ncy
http://www.diabetes.co.uk/insulin/animal-insulin.html
Biology textbook (N.C.E.R.T) Class 12th