Week 6: Genetics (Recombinant DNA and sequences (restriction sites) and cleave the
Genetic Engineering) DNA at these sites, resulting in fragments with
sticky ends.
What is a Recombinant DNA (rDNA)? It is a
type of DNA that is created by combining 3. Inserting DNA: The desired gene or
genetic material from two or more different DNA fragment is inserted into a vector, which
sources. This technique allows scientists to is a carrier molecule that can replicate
artificially manipulate DNA sequences to create independently in a host organism. Common
novel genetic combinations that may not occur vectors include plasmids and viral genomes.
naturally.
4. Joining DNA: The vector and the DNA
The process of creating recombinant DNA fragment are joined together using DNA ligase,
typically involves isolating specific DNA an enzyme that catalyzes the formation of
sequences from one organism, often using phosphodiester bonds between the DNA
enzymes called restriction endonucleases to molecules. This creates a recombinant DNA
cut the DNA at precise locations. These DNA molecule.
fragments can then be combined with the DNA
of another organism, either by inserting them 5. Transformation: The recombinant DNA
into plasmids (small, circular DNA molecules molecule is introduced into a host organism,
found in bacteria) or by directly integrating such as bacteria or yeast, through a process
them into the genome of a host organism. called transformation. The host organism then
replicates the recombinant DNA and produces
Once the recombinant DNA is introduced into a the desired protein encoded by the inserted
host organism, such as bacteria, yeast, or plant gene.
cells, it can be replicated and expressed,
leading to the production of proteins encoded Applications of Recombinant DNA
by the inserted DNA sequences. This ability to Technology:
introduce new genes into organisms and Recombinant DNA technology has numerous
control their expression has numerous applications across various fields:
applications in biotechnology, including the
production of pharmaceuticals, the 1. Medicine: Genetic engineering is used
development of genetically modified crops, and to produce therapeutic proteins, such as insulin
the study of gene function and regulation. and growth hormones, for the treatment of
diseases like diabetes and growth disorders. It
Principles of Recombinant DNA also plays a crucial role in gene therapy, where
Technology: defective genes are replaced or modified to
Recombinant DNA technology involves several treat genetic disorders.
key principles:
2. Agriculture: Genetically modified (GM)
1. Isolation of DNA: The first step in crops are engineered to exhibit desirable traits,
genetic engineering is to isolate the DNA of such as pest resistance, herbicide tolerance,
interest from the organism. This can be and improved nutritional content. These crops
achieved using various techniques, such as help increase yields, reduce pesticide use, and
cell lysis and DNA extraction kits. enhance food security.
2. Cutting DNA: Once isolated, the DNA 3. Industry: Recombinant DNA technology
is cut into fragments using restriction enzymes. is utilized in the production of enzymes,
These enzymes recognize specific DNA biofuels, and bioplastics. Microorganisms like
bacteria and yeast are genetically engineered using ligase enzymes to create a
to produce valuable compounds on an recombinant DNA molecule. This
industrial scale. recombinant DNA is introduced into a host
organism, such as bacteria or yeast, where
4. Research: Genetic engineering it replicates along with the host's DNA,
facilitates the study of gene function, producing multiple copies of the gene.
regulation, and expression. Techniques such Gene cloning has numerous applications,
as gene knockout and gene editing allow including the production of therapeutic
scientists to manipulate specific genes in proteins, creation of genetically modified
model organisms to understand their roles in crops, and study of gene function through
biological processes. gene knockout experiments.
Genetic engineering (also called genetic 2. Polymerase Chain Reaction (PCR): PCR is
modification) is a process that uses a powerful technique used to amplify
laboratory-based technologies to alter the DNA specific regions of DNA, enabling the
makeup of an organism. This may involve production of millions of copies of a target
changing a single base pair (A-T or C-G), DNA sequence. The process involves three
deleting a region of DNA or adding a new main steps: denaturation, where the DNA
segment of DNA. For example, genetic strands are separated by heating;
engineering may involve adding a gene from annealing, where short DNA primers bind
one species to an organism from a different to complementary sequences flanking the
species to produce a desired trait. Used in target region; and extension, where DNA
research and industry, genetic engineering has polymerase synthesizes new DNA strands
been applied to the production of cancer using the primers as templates. PCR is
therapies, brewing yeasts, genetically modified widely used in various fields, including
plants and livestock, and more. molecular biology research, forensic
science, and medical diagnostics, due to its
At its core, genetic engineering involves the sensitivity, specificity, and speed.
deliberate alteration of an organism's genetic
material using biotechnology tools. This 3. Gene Editing: Gene editing technologies
alteration can encompass the addition, allow precise modification of an organism's
deletion, or modification of specific genes, genome by making targeted changes to its
allowing scientists to bestow desired traits or DNA sequence. One of the most prominent
characteristics onto organisms. Techniques gene editing tools is CRISPR-Cas9, which
such as gene splicing, CRISPR-Cas9 gene utilizes a guide RNA molecule to direct the
editing, and recombinant DNA technology Cas9 enzyme to a specific DNA sequence,
serve as the backbone of genetic engineering. where it induces a double-strand break.
The cell's natural DNA repair mechanisms
TECHNIQUES IN GENETIC ENGINEERING: then repair the break, often resulting in
1. Gene Cloning: Gene cloning is a gene knockout, insertion, or modification.
fundamental technique in genetic CRISPR-Cas9 has revolutionized genetic
engineering that involves making copies of engineering by offering a simple, efficient,
a specific gene or DNA sequence. The and versatile method for genome editing in
process typically begins with the isolation a wide range of organisms, including
of the gene of interest using restriction humans. It holds promise for treating
enzymes, which cut DNA at specific genetic disorders, engineering disease-
sequences. The gene is then inserted into resistant crops, and advancing basic
a vector, such as a plasmid or viral DNA, research in genetics.
● Disease Resistance: Genetic
4. Transgenesis: Transgenesis involves the
introduction of foreign genes into an engineering facilitates the production of
organism's genome, resulting in the disease-resistant plants and animals,
expression of novel traits. This can be reducing the need for antibiotics and
achieved using various methods, such as chemical treatments. This can mitigate
microinjection, where DNA is directly the spread of infectious diseases and
injected into fertilized eggs; biolistics, improve animal welfare in agriculture.
where DNA-coated particles are shot into ● Pharmaceutical Production: Genetically
cells using a gene gun; or viral vectors,
engineered microorganisms are used to
where viruses are used to deliver genes
produce valuable pharmaceuticals,
into host cells. Transgenic organisms, such
including insulin, vaccines, and growth
as genetically modified plants and animals,
factors. This scalable and cost-effective
have been developed for agricultural,
approach ensures a stable supply of
pharmaceutical, and research purposes.
essential medicines and reduces
They have been engineered to exhibit traits
dependency on traditional
such as herbicide resistance, pest
manufacturing methods.
resistance, increased nutritional value, and
production of therapeutic proteins. ● Gene Therapy: Genetic engineering
holds promise for treating genetic
Pros of Genetic Engineering: disorders by correcting defective genes
1. Agricultural Advancements: or introducing therapeutic genes into
● Improved Crop Yield: Genetic patients' cells. This personalized
approach to medicine has the potential
engineering enables the development of
to cure previously incurable diseases
crops with desirable traits, such as
and improve patients' quality of life.
resistance to pests, diseases, and
environmental stressors. This can lead
3. Environmental Benefits:
to increased agricultural productivity and
food security, particularly in regions ● Bioremediation: Genetically engineered
prone to crop failures. microorganisms can be deployed for
● Enhanced Nutritional Value: Scientists environmental cleanup, breaking down
pollutants and contaminants in soil,
can engineer crops to have higher
water, and air. This approach offers a
nutritional content, such as increased
sustainable solution to remediate
levels of vitamins, minerals, and
polluted sites and restore ecosystems.
essential nutrients. This has the
potential to address malnutrition and ● Conservation Efforts: Genetic
improve public health outcomes. engineering techniques, such as cloning
● Sustainable Agriculture: By engineering and assisted reproduction, contribute to
species conservation and biodiversity
crops that require fewer pesticides and
conservation efforts. Endangered
fertilizers, genetic engineering can
species can be preserved and
promote sustainable farming practices,
reintroduced into their natural habitats,
reducing environmental pollution and
preventing extinction and restoring
preserving biodiversity.
ecological balance.
2. Medical Breakthroughs:
Cons of Genetic Engineering:
1. Ethical Concerns: effectiveness of antibiotics in treating
bacterial infections.
● Playing "God": Critics argue that genetic
engineering involves tampering with 3. Environmental Risks:
nature and arrogating powers
traditionally attributed to a higher ● Ecological Disruption: Genetically
authority. The notion of manipulating the modified crops may escape cultivation
fundamental building blocks of life raises and interbreed with wild relatives,
ethical questions about the sanctity of leading to unintended consequences
nature and the limits of human such as the spread of transgenes and
intervention. the disruption of native ecosystems.
Concerns exist about the potential for
● Genetic Discrimination: Genetic
genetic pollution and the loss of
engineering may exacerbate existing biodiversity.
social inequalities and lead to
discrimination based on genetic traits. ● Herbicide Resistance: The widespread
Concerns arise regarding access to adoption of genetically engineered crops
genetic enhancements, genetic with herbicide-resistant traits has led to
screening, and the potential for the emergence of herbicide-resistant
eugenics-like practices. weeds, necessitating increased
herbicide use and posing ecological
● Animal Welfare: The genetic
challenges in agricultural landscapes.
modification of animals for agricultural
and research purposes raises ethical
dilemmas related to animal welfare,
suffering, and the intrinsic value of
sentient beings. Questions persist about
the ethical treatment of genetically
engineered animals and the potential for
unintended consequences.
2. Health Risks:
● Unforeseen Side Effects: The long-term
health effects of genetically modified
organisms (GMOs) on human health are
not fully understood. Critics argue that
genetic modifications could introduce
unintended allergens, toxins, or other
harmful substances into the food supply,
posing risks to consumer health.
● Antibiotic Resistance: Genetic
engineering techniques often involve the
use of antibiotic resistance markers to
select for genetically modified cells. This
practice contributes to the spread of
antibiotic resistance genes in the
environment, compromising the
