Biotechnology and Its Applications

Critical Areas of Biotechnology:

- Providing improved organisms or pure enzymes as catalysts.

- Engineering optimal conditions for catalyst action.

- Downstream processing for protein/organic compound purification.

Biotechnological Applications in Agriculture:

- Genetically Modified Organisms (GMOs) are organisms with altered genes.

- Advantages of Genetic Modification in plants:

- Increased tolerance to abiotic stresses (cold, drought, salt, heat)

- Reduced reliance on chemical pesticides (pest-resistant crops)
- Decreased post-harvest losses
- Enhanced mineral usage efficiency by plants
- Improved nutritional value (e.g., vitamin A-enriched rice)

Bt Cotton:

- Bacillus thuringiensis (Bt) produces

proteins toxic to certain insects.

- Bt toxin genes transferred to cotton,

producing insect-resistant plants.

- Toxin activated in insect gut, causing cell

swelling and death.

Pest-Resistant Plants:

- RNA interference (RNAi) used to prevent nematode infestation in tobacco plants.

- Double-stranded RNA (dsRNA) directs degradation of specific mRNA.

Steps of RNA Interference:

1. Production of Double-stranded RNA (dsRNA):

- dsRNA is generated either endogenously or exogenously.

2. Introduction of Nematode-specific Genes into Host Plant:

- Agrobacterium vectors are used to introduce nematode-specific genes into the host plant (e.g., tobacco plant).

3. Production of Sense and Antisense RNA:

- DNA introduction in the host plant leads to the production of both sense and antisense RNA.

4. Formation of Double-stranded RNA (dsRNA):

- Sense and antisense RNA, being complementary, form dsRNA, initiating RNA interference (RNAi).

5. Incorporation of dsRNA into RISC:

- dsRNAs are cleaved into small segments (siRNAs) by the enzyme Dicer.

- siRNAs are then incorporated into the RNA-induced silencing complex (RISC).
6. Targeting mRNA for Degradation:

- Guided by base complementarity of siRNA, RISC targets specific mRNA for degradation.

7. Parasite Elimination:

- The consequence of RNAi is the degradation of nematode mRNA, leading to the parasite's inability to survive in
the transgenic host.

Biotechnological Applications in Medicine:

1. Mass Production of Therapeutic Drugs:

- Biotechnology allows the mass production of safe and effective therapeutic drugs.

- Recombinant therapeutics do not induce unwanted immunological responses.

2. Genetically Engineered Insulin:

- DNA sequences corresponding to human insulin chains A and B are introduced

into E. coli plasmids to produce insulin chains.

- Chain A and chain B are produced separately, extracted, and combined to form
mature human insulin.

3. Gene Therapy:

- Gene therapy aims to cure hereditary or genetic diseases by inserting genes into a person's cells and tissues.

- The first clinical gene therapy was performed in 1990 for a girl with adenosine deaminase (ADA) deficiency.

- ADA deficiency leads to the accumulation of toxic deoxyadenosine, causing damage to immune cells.

Treatment:

1. Bone marrow transplantation: This procedure involves replacing the faulty bone marrow with healthy donor bone
marrow, which can produce functional ADA enzyme.

2. Enzyme Replacement Therapy (ERT): Regular injections of the ADA enzyme are administered to the patient to
supplement the deficient enzyme.

3. Gene Therapy: Lymphocytes from the patient's blood are cultured, and functional ADA cDNA is introduced into
these cells. The genetically modified lymphocytes are then returned to the patient. However, periodic infusions are
required since these cells are not immortal.

4. Early Embryonic Gene Therapy: Introducing functional ADA cDNA into cells at early embryonic stages could
provide a permanent cure.

Molecular Diagnosis:

Early detection of disease is not possible by conventional methods (serum and urine analysis)

1. Recombinant DNA technology.

2. Polymerase Chain Reaction (PCR): PCR can amplify and detect very low concentrations of bacteria or viruses,
making it useful for early diagnosis of infections and genetic disorders.

3. Enzyme-Linked Immunosorbent Assay (ELISA): ELISA is used to detect specific antigens or antibodies, aiding in the
diagnosis of various diseases.
Transgenic Animals:
- Transgenic animals are those that have foreign DNA inserted into them, allowing the expression of the alien genes.

Reasons for Creating Transgenic Animals:

1. Normal Physiology and Development:

- Transgenic animals help study how genes are regulated, their effects on normal body functions, and their impact
on growth and development (e.g., insulin-like growth factor).

2. Study of Diseases:

- Transgenic animals are designed to understand the role of genes in diseases such as cancer, cystic fibrosis,
rheumatoid arthritis, and Alzheimer's.

3. Biological Products:

- Transgenic animals are used to produce human-interest biological products like alpha-1-antitrypsin (for
emphysema), proteins for treating PKU and cystic fibrosis, and Rosie, the transgenic cow that produces human
protein-enriched milk.

4. Vaccine Safety:

- Transgenic mice are developed to test the safety of vaccines before human use, such as testing polio vaccines in
mice.

5. Chemical Safety Testing:

- Transgenic animals are used in toxicity/safety testing to assess the effects of toxic chemicals.

Ethical Issues:
- The Genetic Engineering Approval Committee (GEAC) in India makes decisions about the validity and safety of GM
research and the introduction of GM organisms for public services.

- Biopatents are granted by governments to inventors to prevent others from commercial use of their biological
inventions or products derived from them.

- Biopiracy refers to the unauthorized use or patenting of biological resources by multinational companies and
organizations without proper authorization from the countries and people concerned, without compensatory
payment.