HCS – SCIENCE & TECHNOLOGY

LECTURE- 4 BIOTECHNOLOGY
Biotechnology
• Biotechnology is basically the use of biological systems (microbes, plants and animals) and the
technique to produce substances which are beneficial to people. It includes the manufacture of
antibiotics, Vitamins, vaccines, plastics, disposal of toxic waste using bacteria, pollution control and
production of new fuels, etc. Various techniques deployed in biotechnology are genetic engineering
or recombinant DNA, technology, hybridism technology, cell and tissue culture and protoplasm
fusion. Others can be germplasm development, embryo transfer technology, enzyme and protein
engineering, fermentation, bioconversion and immobilisation of cells and cellular products. On the
basis of the level at which the transfer is made, biotechnology can be classified into two types gene
biotechnology involving gene transfer and non-gene biotechnology involving cells, tissue or the
whole organism.
Department of Biotechnology (DBT)
• In 1986, the Department of Biotechnology (DBT) was established under the Ministry of Science and
Technology to promote the development of the field of modern biology and biotechnology in the
country.
• In order to promote biotechnology, several steps have been taken by DBT that include technological
transfer to industries, patenting of innovations, interactions with thousands of scientists thereby
utilising the existing expertise of the universities and other national laboratories. This can be
considered as a modest beginning. Recently, efforts are being made for transgenic research in plants
with emphasis on pest and disease resistance, nutritional quality, silk-worm genome analysis,
molecular biology of human genetic disorders, brain research, plant genome research, development,
validation and commercialisation of diagnostic kits and vaccines for communicable diseases, food
biotechnology, biodiversity conservation and bioprospecting, setting up of micro-propagation parks
and biotechnology-based development for SC/ST, rural areas and women in different states.
Advisory Committees
• The two apex level committees functioning with the department are advisory in nature. The
committees suggest new areas, assist in priority setting and review monitoring of large inter-
institutional, inter-disciplinary projects.
▪ Scientific Advisory Committee of DBT (SAC-DBT)
▪ Standing Advisory Committee Overseas (SAC-O)
SCOPE OF BIOTECH/BIOTECH IN INDIA
• Blue Biotechnology describes the applications of biotechnology in the marine and aquatic field.
• Green Biotechnology represents application of biotechnology in agricultural processes. Like
selection and domestication of plants via micro propagation, designing and then growing transgenic
plants under specific environments (cold, hot, dry, humid) in the presence (or absence) of chemicals.
It might produce more eco-friendly solutions than traditional industrial agriculture.
• Red Biotechnology applies to medical processes, e.g., designing of organisms to produce antibiotics,
and genetic cures through genetic manipulation.
• White biotechnology, or industrial biotechnology, is the biotechnology used in industrial processes,
which includes designing of an organism to produce a useful chemical. Using of enzymes for
industrial catalysts to either produce valuable chemicals or destroy hazardous/polluting chemicals.
Bio economy represents investment and economic output of all these types of applied
biotechnologies.
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 STEM CELLS
• A stem cell is essentially the building block of the human body.
• The biological cells found in all multicellular organisms capable of dividing (through mitosis) and
differentiate into diverse specialised cell types with self-renewal ability to produce more stem cells
are called stem cells.
• Unique Properties of All Stem Cells:
• Three general properties:
▪ Self-renewal
▪ They are unspecialised
▪ Potency

• As per their potential to differentiate into other types of cells, they can be categorised. Embryonic
stem cells are the most potent since they lead to every type of cell in the body.
▪ Totipotent
▪ Pluripotent
▪ Multipoten
▪ Oligopotent
▪ Unipotent
• Totipotent: These stem cells can be differentiated into embryonic and extra-embryonic cell types,
and thus can construct a complete viable organism.
• Pluripotent: The descendants of totipotent cells are stem cells and can differentiate into nearly all
cells, i.e., cells derived from any of the three germ layers.
▪ Embryonic stem cells come under this category.
▪ Embryonic stem cells are considered pluripotent and not totipotent due to lack of the ability to
become part of the extra-embryonic membranes or the placenta.

• Multipotent: This type of stem cells are differentiated into a number of cells.
▪ Adult haematopoietic stem cells are multipotent.
▪ Adipose tissue is a source of multipotent stem cells
▪ Multipotent cells from multiple blood cell lineages.
Oligo potent
• The corneal epithelium is a squamous epithelium that is constantly renewing and is oligopotent.
UniPotent
• These cells can differentiate into only one cell type, but have the property of self-renewal,
distinguishing them from non-stem cells.
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 Types of Stem Cells
• Embryonic stem cells (ES cells)
▪ Pluripotency : ES cells are pluripotent, which is due to their ability to differentiate into all the
derivatives of the three primary germ layers, namely, ectoderm, endoderm and mesoderm.
▪ Ability to Replicate Indefinitely: Embryonic stem cells can propagate themselves indefinitely
under defined conditions.
• Foetal Stem Cells: They are primitive cell types found in the organs of foetus.
• Adult Stem Cells: Undifferentiated cells, found throughout the body after embryonic development,
which can multiply by cell division to replenish dying cells and regenerate damaged tissues are all
adult stem cells. They are also called somatic stem cells and are present in juveniles as well as adult
animals and humans.
The use of adult stem cells in research and therapy is not considered to be controversial, since they are
derived from adult tissue samples instead of destroyed human embryos, as in the case of embryonic
stem cells.

• Amniotic Stem Cells: Stem cells can be used from amniotic fluid, the ethical objection to use human
embryos as a source of cells is currently not an issue.
• Induced Pluripotent Stem Cells:
• First, iPSCs were produced in 2006 from mouse cells and in 2007 from human cells. This may allow
researchers to obtain pluripotent stem cells, important in research and potentially having
therapeutic uses, without the controversial use of embryos. Because iPSCs are developed from a
patient’s own somatic cells, it was expected that treatment of iPSCs would avoid any immunogenic
response.
Stem Cell Treatment:
• It is a type of intervention strategy to induce new cells into damaged tissue so as to treat disease or
injury. The self-renewal ability and capability to give rise to subsequent generations with variable
degrees of differentiation capacities offers significant potential to generate tissues that can
potentially replace diseased and damaged areas in the body. The risk of rejection and side effects
will also be minimal in the case.
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 Stem Cell Controversy:
• The stem cell controversy is the ethical debate centred only on research involving the creation, usage
and destruction of human embryos. Most commonly this controversy focuses on embryonic stem
cells.
India’s Top Centres for Stem Cell Research:
• India’s top centres for stem cell research include the following:
▪ Institute for Stem cell Biology and Regenerative Medicine in stem in Bengaluru.
▪ Centre for Stem Cell Research (CSCR), Christian Medical College (CMC), Vellore, Tamil Nadu.

• School for Regenerative Medicine, Manipal, Karnataka.

• Centre for Stem Cell Science, Hyderabad
• International Centre for Stem Cell, Cancer and Bio-technology in Pune.
• National Centre for Cell Science in Pune
• L.V. Prasad Eye Institute, Hyderabad.
Stem cell therapy (SCT)
• is the treatment of various disorders by using stem cells. The stem cells can be procured from a lot
of different sources and used to potentially treat more than approved disorders.
• Hematopoietic disorders (eg. Leukaemia, thalassemia, aplastic anaemia, sickle cell Anaemia, storage
disorders etc.) are being treated by SCT.
• Certain degenerative disorders [due to wear and tear of bone, cartilage, muscle, fat or any other
tissue/organ] such as Diabetes, stroke, osteoarthritis, chronic renal failure, Alzheimer’s disease,
Parkinson’s disease etc possess a potential treatment by SCT.
STEM CELLSCAN BE OBTAINED FROM DIFFERENT SOURCES:
• Autologous,: From Patient himself
• Syngeneic,;From Patient himself
• allogenic:related/unrelated or cord blood donor
PROCESS OF SCT:
• Obtaining stem cells from a patient or a donor blood stem cells (apheresis). Blood is taken from a
vein and circulated through a machine that removes the stem cell and returns remaining blood and
plasma back to the patient.
• Bone marrow stem cells are harvested from donors by placing a needle in soft marrow of bone.
• Conditioning Treatment : Administration of chemotherapy/radiation therapy to the patient in order
to destroy all diseased cells in the body and to create space in bone marrow for transplanted stem
cells to populate.
• Infusion of healthy stem cells to the patient through intravenous infusion.
• Engraftment and recovery: Generally takes 2-3 weeks. The patient stays in the hospital during this
period and monitored for side effects/infections etc.
Advantage of SCT over conventional treatments:
• Owing to amazing and unique features of stem cells, SCT has been showing a lot of promising results
all over the world.
• The treatment is safe, speedy and the results are for better than long and ineffective conventional
therapies.
• iPSCs (e.g. Cells made from someone’s own organ/skin) have a great advantage since they are
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unlikely to be rejected by the body’s immune system.

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 • Cord blood stem cells and amniotic fluid stem cells harvested after child birth can be frozen in cell
banks and can be used to treat children with blood cancers and genetic blood disorders.
• Using stem cells to test new drugs. Researchers are also using differentiated stem cells to test the
safety and effectiveness of new medications. Testing drugs on human stem cells eliminates the need
to test them on animals.

GENETIC ENGINEERING
• Genetic engineering is a technique of manipulating the genome of the organism using biotechnology
to add one or more traits that are not found in the organism naturally. The technique is also called
gene manipulation/genetic modification.
• Several important products of medical use such as insulin and the human growth hormone are now
being produced commercially by modifying the genome of the bacteria.
Recombinant DNA Technology (RDT)
• Recombinant DNA technology is the definite protocol or method deployed to introduce foreign DNA
segments (DNA of other species) into host species. Even the process followed in gene cloning
(described in the cloning section) is also called recombinant DNA technology. Recombinant DNA
represents DNA from a foreign source.
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 • Isolation of Genes Desirable sequence of genes is obtained directly from the genome of normal cell
or from other cells, which is achieved by cleavage and denaturation. DNA is extracted from the cells.
• Synthesis of Genes Various methods are developed for gene synthesis.
• Recombinant DNA Cutting of DNA molecule at a desired position results in a new gene product which
is called as recombinant DNA (r-DNA). The receiving organism is said to be transgenic. Using this
technique, we can isolate and clone single copy of a gene or a DNA molecule into an indefinite
number of copies, all identical.
• Gene Cloning Isolation of gene and reproduction of a single copy of gene or DNA segment into an
infinite number of copies, all identical, is known as gene cloning. The vectors such as plasmids and
phages reproduce in their usual style even after insertion of foreign DNA. The inserted DNA will also
replicate with the parent DNA. Recently extensive use of newly discovered polymerase chain
reaction (PCR) has also been made for gene technology.
Hazards of Genetic Engineering
• If a wrong DNA segment is inserted and if it gets expressed, it can cause new diseases in human
beings.
• It can be used in biological warfare.
• Genetic modification of existing species/recreation of extinct species can cause disaster.
• Scientists are prone to making mistakes – new strains of bacteria, fauna, etc., can come out of labs
which can be hostile to human beings.
• Even in a single species, genetic engineering leads to elimination of varieties – if some new disease
comes up, the entire species may be wiped out.
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 Applications of Recombinant DNA Technology rDNA Technology:
• Even biological technology, western pharmacy, doctor’s or veterinarian’s office, medical testing
laboratory and biological research laboratory using this technology, deploys application of this
technology, one way or the other.
Some specific examples are as follows:
• Recombinant human insulin: Insulin dependent diabetes has seen a major breakthrough in its
treatment with the production of recombinant insulin. It is cheaper and easily available as compared
to insulin obtained from animal sources.

• Recombinant human growth hormone (hGH, somatotropin): For patients with pituitary glands
generating insufficient quantity of hormone for normal growth and development, it proves to be a
boon. Earlier, when it was not available, HGH was obtained from pituitary glands of cadavers.
Recombinant HGH solved this issue, and is now used therapeutically. Some athletes have been
found misusing it as a performance-enhancing drug.
• Recombinant blood clotting factor VIII: A blood-clotting protein that helps in curing patients with
bleeding disorder haemophilia which is the inability to produce factor VIII in sufficient amount that
can support normal blood coagulation.
• Recombinant hepatitis B vaccine: Recombinant hepatitis B vaccine can control infection of hepatitis
B which carries a form of the hepatitis B virus surface antigen, produced in yeast cells. Hepatitis B
virus, unlike other common viruses such as polio virus, cannot be grown in vitro; it marks to be an
important and necessary development.
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 • b-carotene (pro-vitamin A) is produced in green tissues of rice plants and not in the endosperm or
edible part of the seed. The aleurone layer of husked grain contains a number of nutrients, such as
vitamin B and nutritious fats, but lack pro-vitamin A.
• Golden rice: It is a recombinant variety of rice expressing the enzymes responsible for b-carotene
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biosynthesis. It is advantageous being capable of reducing the deficiency of vitamin A in the world’s
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population.

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 • Golden rice is not currently in use due to pending resolution of intellectual property, environmental
and nutritional issues.
Herbicide and Insect-resistant crops
• Commercial varieties of important agricultural crops (like soya, maize/corn, sorghum, canola and
cotton) have been developed. Such verities integrate a recombinant gene that cause the resistance
to the herbicide glyphosate (trade name Roundup), and also simplifies weed control by glyphosate
application.
Bt Crops
The Belgian company Plant Genetic Systems emerged as the first company (in 1985) aimed to develop
genetically engineered tobacco plants with insect tolerance by expression of cry genes from Bt.
• Bt stands for Bacillus thuringiensis, a bacteria that lives in soil and is commonly used as a biological
alternative to a pesticide; a trans gene producing Cry toxin is extracted by B. thuringiensis and
introduced in desirable plant and develops genetically engineered plants variety such as Bt crop.
Some examples include varieties such as Bt cotton that protect them from insect named Lepidoptera
(Boll worm).
• The first and only transgenic crop approved by the “Genetic Engineering Approval committee”
(GEAC) for commercial cultivation in six States, namely, Andhra Pradesh, Gujarat, Karnataka,
Madhya Pradesh, Maharashtra and Tamil Nadu is named the Bt Cotton.

• Genetically Modified organism or genetically engineered organism (GEO) is an organism with

altered, genetic material due to DNA / genetic engineering technology. So, the organism so
developed has the desirable DNA segment from different sources, combined into one molecule. And
this creates a new set of genes. Thus modifying the genome of the organism
• Transgenesis: this process by which an exogenous or external gene also called as a transgene is
inserted into an organism is called Transgenesis.
Transgenic animals
• They serve as experimental models to carry out phenotypic and other biomedical research. GMO
development is very useful in cures and treatments of many serious diseases. By changing the nature
of DNA ((DNA base pair sequences) on insertion of other DNA or gene to an animal (Sheep, pigs and
rats), certain useful medical treatments can be possible. For instance, Human alpha - I- antitrypsin,
developed in sheep, treats humans with this deficiency, and transgenic pigs with human
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histocompatibility have been situated with primary objective to check will be Suitable for “transplant
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organs will be suitable" for' transplant with low chances of rejection.

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 Hazards of Genetic Engineering
• If a wrong DNA segment is inserted and if it gets expressed, it can cause new diseases in human
beings.
• It can be used in biological warfare.
• Genetic modification of existing species/recreation of extinct species can cause disaster.
• Scientists are prone to making mistakes – new strains of bacteria, fauna, etc., can come out of labs
which can be hostile to human beings.
• Even in a single species, genetic engineering leads to elimination of varieties – if some new disease
comes up, the entire species may be wiped out.
DNA Fingerprinting or Genomic Fingerprinting
• DNA fingerprint is a characteristic feature of every individual. It is a technology which is adopted by
the forensic scientists to identify the individuals by their respective DNA profiles. It is also called as
DNA profiling, DNA testing, DNA typing, or genetic fingerprinting or gene mapping or genography.
Basis of DNA Fingerprinting
• The chemical structure of the DNA of every individual is the same but the difference is in the order
of the base pairs. The information contained in DNA is determined primarily by the sequence of
letters along the zipper.
• 99.9% of human DNA sequences are similar in every person, only 0.1% of human DNA is unique.
• Amongst the genome of the individual, are dispersed VNTR’s t.e. Variable number of tandem repeats
(also called minisatellites). Each repeating unit comprises a sequence of 16-64 base pairs.
• By using restriction enzymes specific for sites flanking the VNTR, fragments of variable lengths in
different individuals can be obtained because each individual has a different number of repeats.
• Except for identical twins, it is highly unlikely that 2 individuals with the same pattern will be found
because it is an individual characteristic.
• These minisatellites can be analysed by Southern Blotting, to give a characteristic banding pattern.
• These minisatellites have Mendelian Inheritance so that some bands detected in an individual will
be common with those of mother and other with those of father.
DNA Fingerprinting is performed in the following research institutes in India
• Centre for cellular and molecular biology (CCMB), Hyderabad.
• Centre for DNA fingerprinting (CDFD), Hyderabad.
• National Bureau of Plant Genetic resource (NBPGR), New Delhi.
• National Institute of Plant and Genetic research (NIPGR), New Delhi
Process of DNA Fingerprinting
It includes the following steps:

• DNA Isolation
• Cutting, sizing, and sorting. Specific enzymes known as restriction enzymes (MOLECULAR
SCISSORS)are used for cutting of the DNA at specific places.
• Transfer of DNA to nylon. The distribution of DNA pieces is transferred to a nylon sheet by placing
the sheet on the gel and soaking them overnight.(GEL ELECTROPHORESIS)
• Probing. When radioactive or coloured probes are added to the nylon sheet, it produces a pattern
called the DNA fingerprint.
• DNA fingerprint: The final DNA fingerprint is built by using several probes (5-10 or more) at the same
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time.
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• Blotting Techniques used in different segment measurement are as follows:

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 • Southern Blotting - DNA
• Northern Blotting - DNA
• Western Blotting - Protein

Practical Applications of DNA Fingerprinting

• Paternity and Maternity Analysis of VNTR’s pattern of Parent-child has been used to solve the father-
identification cases.

• In Forensic sciences DNA isolated from blood, hair, skin cells or other genetic evidence left at the
scene of a crime can be used to identify criminals. Police all over the world are using this technique
to link the suspects with their biological evidence.
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 • Personal Identification DNA fingerprints can be used as genetic barcode to identify individuals.
• Diagnosis of Inherited Disorders It can help to diagnose inherited disorders in both prenatal and new-
born babies. Disorders included under these may include cystic fibrosis, haemophilia, Huntington’s
disease, familial Alzheimer’s, sickle cell anaemia, thalassemia and many others.
DNA Technology (Use and Application) Regulation Bill, 2018
• According to the draft report of the Parliamentary Standing Committee on Science and Technology,
the DNA Technology (Use and Application) Regulation Bill, 2018 could be misused for caste or
community-based profiling.
Key Points
Key Provisions:
• The Bill allows the use of the technology to establish the identity of persons in matters of crime,
parentage dispute, emigration or immigration and transplantation of human organs.
• It provides for establishment of national and regional DNA data banks and each databank will
maintain crime scene index, suspects’ or undertrials’ index and offenders’ index separately.
Background:
• A similar bill was passed in Lok Sabha in 2018 but could not be passed in the Rajya Sabha.
• The 2019 Bill was referred to the Parliamentary Standing Committee on Science and Technology,
Environment and Forests for examination.
Concerns Raised by the Committee:

• Misuse of Sensitive Information:

▪ The DNA profiles can reveal extremely sensitive information of an individual such as family
ancestry (pedigree), skin colour, behaviour, illness, health status and susceptibility to diseases.
▪ Access to such intrusive information can be misused to specifically target individuals and their
families with their own genetic data.
▪ It could even be used to incorrectly link a particular caste/community to criminal activities.
▪ Storage of DNA Profiles of Unconvicted Persons:

▪ The Bill proposes to store DNA profiles of suspects, undertrials, victims and their relatives for
future investigations.
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▪ The Bill also provides that DNA profiles for civil matters will also be stored in the data banks, but
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without a clear and separate index.

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 ▪ The committee has questioned the necessity for storage of such DNA profiles, pointing out that
this violates the fundamental right to privacy and does not serve any public purpose.
• Perfunctory Consent:

▪ The Bill refers to consent in several provisions, but in each of those, a magistrate can easily
override consent, thereby in effect, making consent perfunctory.
▪ There is also no guidance in the Bill on the grounds and reasons of when the magistrate can
override consent.

• Removal of DNA Profiles of Accused:

▪ The Bill permits retention of DNA found at a crime scene in perpetuity, even if conviction of the
offender has been overturned.
▪ The committee has recommended that independent scrutiny must be done of the proposals to
destroy biological samples and remove DNA profiles from the database.

• Absence of Robust Data Protections

▪ The committee has also called the Bill “premature” and questioned the security of a huge
number of DNA profiles that will be placed with the National DNA Data bank and its regional
centres.
• Need of the Bill:

▪ DNA testing is currently being done on an extremely limited scale in India, with approximately
30-40 DNA experts in 15-18 laboratories undertaking less than 3,000 cases per year, which
represent 2-3% of the total need.
▪ The standards of the DNA testing laboratories are not monitored or regulated, in absence of any
proper regulation.
▪ The Bill will enable identification of missing children.
▪ As per the National Crime Records Bureau, annually 1,00,000 children go missing.
o The Bill will also help in identifying unidentified deceased, including disaster victims and
apprehend repeat offenders for heinous crimes such as rape and murder.
o Over the concerns of misuse it is being said that any and every blood sample collected by a
clinical laboratory has the same potential for misuse.
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