Biotechnology and its Applications

Biotechnological Applications in Agriculture

● Genetic modification has:

1. Made crops more tolerant to abiotic stresses (cold, drought, salt, heat)
2. Reduced reliance on chemical pesticides (pest-resistant crops)
3. Helped to reduce post-harvest losses
4. Increased efficiency of mineral usage by plants (this prevents early exhaustion of
the fertility of soil)
5. Enhanced nutritional value of food, e.g., golden rice, i.e., Vitamin A enhanced
rice
● Bt cotton:
a. Some strains of Bacillus thuringiensis produce proteins that kill certain insects
such as lepidopterans (tobacco budworm, armyworm), coleopterans (beetles), and
dipterans (flies, mosquitoes)
b. B. thuringiensis forms protein crystals during a particular phase of their growth,
which contains a toxic insecticidal protein
c. The Bt toxin protein exists as inactive protoxins. Still, once an insect ingests the
inactive toxin, it is converted into an active form of toxin due to the alkaline pH
of the gut, which solubilizes the crystals.
d. The activated toxin binds to the surface of midgut epithelial cells and creates
pores that cause cell swelling and lysis and eventually cause the insect's death.
● Pest-resistant plants:
a. Meloidogyne incognita infects the roots of tobacco plants and causes a significant
reduction in yield
b. A novel strategy was adopted to prevent this infestation which was based on the
process of RNA interference (RNAi)
c. RNA interference (RNAi) - the silencing of a specific mRNA due to a
complementary dsRNA molecule that binds to and prevents translation of the
mRNA (silencing)
● DNA technology enables scientists to produce transgenic animals, which speeds up the
transgenic animals, which speeds up the selective breeding process
● However, health problems are not uncommon among farm animals carrying genes from
other species, and modification of the animal’s genes using the CRISPR-Cas9 system will
likely emerge as a more helpful technique
● Animal health and welfare are important issues to consider when genetically altering
animals
● Genetic manipulations can be performed on an ordinary somatic cell, and the cell then
can be used to generate a plant with new traits
 ● Genetic engineering is rapidly replacing traditional plant-breeding programs, especially
for useful traits, such as herbicide or pest resistance, determined by one or a few genes
● Crops engineered with a bacterial gene making the plants resistant to a herbicide can
grow while weeds are destroyed, and genetically engineered crops that can resist
destructive insects reduce the need for chemical insecticides
● Predictions by the Food and Agriculture Organization of the United Nations:
a. We will need 70% more food by the year 2050 than the planet is currently
producing
b. International C4 Rice Project - the aim of the researchers engaged in this project
is to genetically engineer a strain of rice that can use C4 photosynthesis, which is
more efficient than the C3 form of photosynthesis used by the standard form of
rice
c. In 2017, the first step was accomplished when they were able to engineer a corn
gene into rice plants genetically

Biotechnological Applications in Medicine

● Genetically engineered insulin:

a. Insulin used for diabetes was earlier extracted from the pancreas of slaughtered
cattle and pigs; causing some patients to develop an allergy or other types of
reactions to the foreign protein
b. Insulin consists of two short polypeptide chains - chain A and chain B, that are
linked together by disulfide bridges
c. Insulin prohormone contains an extra stretch of a peptide called the C peptide
d. C peptide is not present in the mature insulin and is removed during maturation
into insulin
e. 1983 - Eli Lilly - an American company - prepared two DNA sequences
corresponding to A and B, chains of human insulin, and introduced them in the
plasmids of E. coli to produce insulin chains; chains A and B were produced
separately, extracted, and combined by creating disulfide bonds to form human
insulin
● imatinib (trade name Gleevec) is a small molecule that inhibits one tyrosine kinase
● The overexpression of tyrosinase kinase resulting from a chromosomal translocation is
instrumental in causing chronic myelogenous leukemia (CML)
● Patients in the early stages of CML who are treated with imatinib have exhibited nearly
complete, sustained remission from the cancer
● Specific tumor cells have a random mutation that allows them to survive in the presence
of a particular drug, and as a consequence of natural selection in the presence of the drug,
these are the cells that survive and reproduce
● Some 2 million people with diabetes in the United States depend on insulin treatment to
control their disease
● Tissue plasminogen activator (TPA) - administered shortly after a heart attack, TPA
helps dissolve blood clots and reduces the risk of subsequent heart attacks
● Transgene - a gene that has been transferred into one organism from another
● Protein production by “Pharm” Animals:
a. Removing eggs from a female of the recipient species and fertilizing them in vitro
b. A transgene for a human blood protein such as antithrombin, which prevents
blood clots, can be inserted into the genome of a goat in such a way that the
transgene’s product is secreted into the animal milk
● Forensic Evidence and Genetic Profiles:
a. If enough blood, semen, or tissue is available, forensic laboratories can determine
the blood type or tissue type by using antibodies to detect specific cell-surface
proteins
b. However, such tests require fairly fresh samples in relatively large amounts; also,
because many people have the same blood or tissue type, this approach can only
exclude a suspect; it cannot provide strong evidence of guilt.
c. DNA testing, on the other hand, can identify the guilty individual with a high
degree of certainty because the DNA sequence of every person is unique (except
for identical twins)
d. Genetic profile - the individual’s unique set of genetic markers
e. Genetic profiles can also identify victims of mass casualties; the most significant
such effort occurred after the attack on the World Trade Center in 2001; more than
10,000 samples of victims’ remains were compared with DNA samples from
personal items, such as toothbrushes, provided by families
f. Ultimately, forensic scientists succeeded in identifying almost 3,000 victims using
these methods
 Gleevec is an effective medicine for treating CML at its early stages.

[Picture credit: Gleevec - Cancer Health]

Human Gene Therapy and Gene Editing

● Gene therapy - the introduction of genes into an afflicted individual for therapeutic
purposes - holds great potential for treating the relatively small number of disorders
traceable to a single defective gene
● The aim of this approach - is to insert a normal allele of the defective gene into the
somatic cells of the tissue affected by the disorder
● A more direct approach that avoids the complications of using a viral vector in gene
therapy is made possible by gene editing
● Technical issues:
a. Controlling the activity of the transferred gene so that cells make appropriate
amounts of the gene product at the right time and in the right place
b. Being sure that the insertion of the therapeutic gene does not harm some other
necessary cell function
● Gene therapy and gene editing provoke ethical questions; some critics say that tampering
with human genes is in any way unethical
● Issue of engineering human germ-line cells to try to correct a defect in future generations
● The development of the CRISPR-Cas9 system has engendered much debate about the
ethics of gene editing
● Jennifer Doudna - co-discoverer of CRISPR-Cas9 - et al. agreed to exercise extreme
caution as the field moves forward
● They called for the research community to “strongly discourage” any experimental work
on human eggs and embryos
● Case of the Chinese Researcher:
a. In 2019, he reported 2018 using the CRISPR-Cas9 system to edit genes in
embryos that completed fetal development and were born as twins and a third
individual
b. He claimed to have edited the CCR5 gene, which codes for a co-receptor for HIV,
so HIV would be unable to bind and infect the cells.
c. The twins’ father was HIV-positive, which was the rationale the scientist used for
this genetic engineering.
d. The biological community condemned this act as highly unethical; the researcher
lost his job and may face a criminal investigation.
● In 2019, an advisory panel to the World Health Organization proposed establishing a
registry to monitor any gene-editing research in humans.
 Figure showing CCR5 receptor protein on the cell surface along with HIV

[Picture credit: CCR5 - Wikipedia]

Molecular Diagnosis

● Techniques that serve the purpose of early diagnosis:

1. Recombinant DNA Technology
2. Polymerase Chain Reaction (PCR)
3. Enzyme-Linked Immuno-sorbent Assay (ELISA)
● Very low concentration of a bacteria or virus (at a time when the symptoms are not yet
visible) can be detected by amplification of their nucleic acid by PCR
● PCR is now routinely used to detect HIV in suspected AIDS patients and detect
mutations in genes in suspected cancer patients too
● PCR is a powerful technique to identify several other disorders apart from HIV and
cancer
● DNA/RNA Probe method:
1. A single-stranded DNA or RNA, tagged with a radioactive molecule (probe), is
allowed to hybridize with its complementary DNA in a clone of cells, followed by
detection using autoradiography
2. The clone with the mutated gene will not appear on the photographic film because
the probe will not have complementarity with the mutated gene.
● ELISA is based on the principle of antigen-antibody interaction

Uses of transgenic animals

● Normal physiology and development - transgenic animals can be specifically designed to

study how genes are regulated and affect the body's normal functions and evolution.
● Study of disease - many transgenic animals are designed to increase our understanding of
how genes contribute to disease development.
 ● Biological products - transgenic animals that produce useful biological products can be
created by introducing the portion of DNA (or genes) that codes for a particular product,
such as human protein (ɑ-1-antitrypsin) used to treat emphysema.
● Vaccine safety - transgenic mice, are being developed for testing the safety of vaccines
before they are used on humans.
● Chemical safety testing - transgenic animals, are made that carry genes that make them
more sensitive to toxic substances than non-transgenic animals.

Personal Genome Analysis

● Direct-to-consumer genome analysis companies offer kits allowing individuals to send in

a swab containing cheek cells that the company will analyze genetically.
● Genetic testing for risk factors like heart disease, Alzheimer’s disease, and some types of
cancer is carried out by looking for linked SNPs that have been previously identified.
● This may be helpful for individuals to learn about their health risks, understanding that
such genetic tests merely reflect correlations and do not make predictions.
● In addition to health-related genetic information, these companies compare a person’s
DNA segments with those from reference populations around the world, established from
thousands of individuals of known ancestry.

Personalized Medicine

● By analyzing the expression of many genes in large numbers of breast cancer patients,
researchers can refine their understanding of the different subtypes of breast cancer.
● Given that some low-risk patients have a 96% survival rate over ten years with no
treatment, gene expression analysis allows doctors and patients access to valuable
information when considering treatment options.
● Personalized Medicine - a type of medical care in which each person’s specific genetic
profile can provide information for which the person is especially at risk and help make
health-care decisions
● Genetic profile - currently taken to mean a set of genetic markers such as SNPs
● Pharmacogenetics - An approach in which an individual’s genomic information can be
used to predict the benefits and risks of particular medications
● There are over 300 medications for which the Food and Drug Administration
recommends genetic testing for patients before their use.
Personalized medicine has been gaining importance over the past few years. It is a very efficient
method to treat a person without many side effects.

[Picture credit: Personalized Medicine - EULAC PerMed]

Biopiracy

● Refers to the use of bio-resources by multinational companies and other organizations

without proper authorization of the countries and people concerned without
compensatory payment
● In 1997, an American company got patent rights on Basmati Rice through the US Patent
and Trademark Office, which allowed the company to sell a ‘new’ variety of Basmati in
the US and abroad, derived from Indian farmers’ varieties
● The patent extended to functional equivalents, implying that the patent could restrict
other people selling Basmati rice
● Several attempts have also been made to patent uses, products, and processes based on
Indian traditional herbal medicines, e.g., turmeric, neem
● The Indian Parliament has recently cleared the second amendment of the Indian Patents
Bill, which takes issues related to biopiracy under consideration, including patent terms,
emergency provisions, and research and development initiative

Safety and Ethical Questions Raised by DNA Technology

● Early concerns about potential dangers associated with recombinant DNA technology and
genetic engineering focused on the possibility that hazardous new pathogens might be
created
● To guard against such rogue microorganisms, scientists developed a set of guidelines that
were adopted as formal government regulations in the United States and some other
countries
● The Indian Government has set up organizations such as the GEAC (Genetic Engineering
Approval Committee), which will make decisions regarding the validity of GM research
and the safety of introducing GM organisms to public services
● Strict laboratory procedures - designed to prevent engineered microorganisms from
either infecting researchers or accidentally leaving the laboratory
● Strains of microorganisms to be used in recombinant DNA experiments are genetically
crippled to ensure that they cannot survive outside the laboratory
● Finally, specific dangerous experiments have been banned
● Genetically modified organisms (GMO):
a. A transgenic organism that has acquired one or more genes from another species
or another variety of the same species
b. The majority of GMOs that contribute to our food supply are not animals, but
crop plants
c. GM crops are widespread in the United States, Argentina, and Brazil; together,
these countries account for over 80% of the world’s acreage devoted to such
crops.
d. In the United States, most corn, soybean, and canola crops are genetically
modified, and a recent law requires labeling GM crops.
e. In Europe, the European Union established a comprehensive legal framework
regarding GMOs in 2015
f. The high consumer distrust in Europe makes the future of GM crops there
uncertain.
g. Some people fear that the protein products of transgenes might lead to allergic
reactions.
● Today, governments and regulatory agencies worldwide are grappling with how to
facilitate the use of biotechnology in agriculture, industry, and medicine while ensuring
that new products and procedures are safe.
● Advances in biotechnology have allowed us to obtain complete genome sequences for
humans and many other species, providing a vast treasure trove of information about
genes.
● Ethical considerations and concerns about potential environmental and health hazards
will likely slow some applications of biotechnology.
● Given the tremendous power of DNA technology, we must proceed with humility and
caution.
Text References

● Biology Class 12 - NCERT

● Campbell Biology Twelfth Edition
● Introduction to Genetic Analysis 9th Edition
● Genetics Analysis and Principles Fourth Edition