Genetic Engineering

techniques
• Artificial Selection (ancient
time)
• Selective Breeding
• Hybridization
• Inbreeding
• Cloning
• Recombinant DNA
• Polymerase Chain Reaction
(PCR)
• Gel Electrophoresis
• Transgenic Organisms
 Artificial Selection
• Artificial selection is done to indirectly
manipulate genes focusing on the physical traits
among organisms. Breeders choose which
organism to mate and produce offspring with
desirable traits.
 Selective Breeding
• Breed only those
plants or animals with
desirable traits

• People have been

using selective
breeding for 1000’s of
years with farm crops
and domesticated
animals.
 Hybridizations
• Hybridizations are when two
individuals with unlike
characteristics are crossed
to produce the best in both
organisms like the disease
resistant potato called the
Burbank potato.
• Russet Burbank is
a potato cultivar with dark
brown skin and few eyes
that is the most widely
grown potato in North
America.
 Inbreeding
• Inbreeding is a technique of
breeding organisms that are
genetically similar to
maintain desired traits found
in the pure dog breeds
 Cloning
• Cloning is a
technique
scientists use to
make exact
genetic copies
of living things.
Genes, cells,
tissues, and even
whole animals
can all be cloned.
 Cloning
• Cloning provides strong evidence that
differentiated cells retain their full genetic
potential.
• Animal cloning is achieved through a
procedure called nuclear transplantation.
– Involves replacing the nucleus of an egg cell or
zygote with the nucleus of adult somatic cell.
– The egg cell may then begin to divide.
– About 5 days later, repeated cell divisions form
a blastocyst, a ball of cells.
– At this point, the blastocyst may be used for
different purposes.
 Cloning
1. Reproductive cloning
– If the animal to be cloned is a mammal,
further development requires implanting the
blastocyst into the uterus of a surrogate
mother.
– The resulting animal will be genetically
identical to the donor of the nucleus—a
“clone” of the donor.
– This type of cloning results in the birth of a
new individual
 Cloning
2. Therapeutic cloning
– Embryonic stem cells (ES cells) are
harvested from the blastocyst.
– In nature, embryonic stem cells give rise to all
the different kinds of specialized cells of the
body.
– In the laboratory, embryonic stem cells are
easily grown in culture, where, given the right
conditions, they can perpetuate themselves
indefinitely.
 Recombinant DNA
• The ability to combine the
DNA of one organism with the
DNA of another organism.
• Recombinant DNA technology
was first used in the 1970’s
with bacteria.
 Recombinant DNA
 The altered DNA is called recombinant
DNA
 Recombinant DNA is joined to other
unrelated DNA in the organism
 This is called gene splicing.
- tiny segments of a gene are taken out
and replaced by different genes
Recombinant Bacteria
1. Remove bacterial DNA
(plasmid).

2. Cut the Bacterial DNA with

“restriction enzymes”.

3. Cut the DNA from another

organism with “restriction
enzymes”.

4. Combine the cut pieces of DNA

together with another enzyme
and insert them into bacteria.

5. Reproduce the recombinant

bacteria.

6. The foreign genes will be

expressed in the bacteria.
How to Create a Genetically
Modified Plant

1.Create recombinant
bacteria with desired
gene.

2. Allow the bacteria to

“infect" the plant cells.

3. Desired gene is
inserted into plant
chromosomes.
 Polymerase Chain Reaction (PCR)
- allows scientists to make many copies of a
piece of DNA
1. Heat the DNA so it
“unzips”.

2. Add the
complementary
nitrogenous bases.

3. Allow DNA to cool

so the
complementary
strands can “zip”
together.
 PCR
1977
• PCR – Polymerase Chain Reaction
• Kary Mullis
• Idea just came to him while he was
thinking about DNA Replication (on LSD)
• Won Noble Prize
 PCR PROCEDURE
1. Separate the Double Helix
2. Bind primers (2) to sequence you want to
replicate
3. DNA Polymerase copies between two
primers
4. Rinse and Repeat
5. Copies DNA between two primers
exponentially
 PCR
1. Separate the Double Helix

Heat

2. Bind primers (2) to sequence you want to

replicate

Add
 PCR Process
1. Separate the Double Helix

Heat

2. Bind primers (2) to sequence you want to

replicate

Add
 PCR Process
3. DNA Polymerase copies between two
primers
Add
DNA Pol.
4. Repeat Many Times
PCR
Benefits of Genetic
Engineering

Presented by:
Mrs. Carolyn S. Delmindo
 QUESTION
What are the objectives of
scientists in genetic
engineering?
The aim of planting genetically
modified organism and its
commercial use is to help improve
farming methods efficiently and
productively. Some of the most
common crops are soybean, maize,
cotton, canola, alfalfa, papaya,
eggplant, potato, apple, sunflower,
pineapple, and sugarcane.
Genetically modified plants have created
resistance to harmful agents, enhanced
product yield, and shown increased
adaptability for better survival.
Recombinant technology has been
widely used in improving crop varieties.
Several transgenic or genetically
modified organisms (GMO) have been
produced. Genetically Modified organism
are also called transgenic organism,
since genes are transferred from one
organism to another.
• Recombinant DNA technology is playing a vital role in
improving health conditions by developing new vaccines
and pharmaceuticals.
• The treatment strategies are also improved by
developing diagnostic kits, monitoring devices, and new
therapeutic approaches.
• Synthesis of synthetic human insulin and erythropoietin
by genetically modified bacteria; human growth which
cure stunted growth; tissues plasminogen activator
which dissolves blood clots among patients who had
heart attack; transgenic pigs used to produced human
hemoglobin; human clotting factors have also been
produced in the milk of transgenic goats; production of
“pharm” animals to synthesize such as pharmaceutical
products and production of new types of experimental
mutant mice for research purposes.
Benefits of Genetic Engineering
• As defined Genetic engineering is the
processes of changing the DNA in living
organisms to create something new. It
involves artificial manipulation, modification,
and recombination of DNA or other nucleic
acid molecules in order to modify an
organism or population of organisms.
Recombinant DNA technology has
applications various area like medicine
focusing on the discoveries of medicine to
cure and improve human health and nutrition
aimed to produce good quality and high
quantity crops.
 Benefits of Recombinant
Bacteria
1. Bacteria can make human insulin or
human growth hormone.
2. Bacteria can be engineered to “eat” oil
spills.
Production of human insulin in
bacteria
The transfer of human gene to bacteria
Escherichia coli aims to have a mass
production of human insulin to cure
diabetes mellitus. Insulin gene is
extracted from a human cell and spliced
into a plasmid vector before inserted into
a bacterial cell. The transgenic bacteria
are selected and cultured to be able to
increase its number carrying human
insulin gene. The bacteria are harvested,
purified and packed for human use.
List of genetically modified organisms
Activity 1. Answer the puzzle by writing your answers
in the grid.
 Activity 2 Transgenic (GM) Organisms
Directions. Name the transgenic organism based from the description given
by filling up the missing letters.
 Activity 3 Uses of Transgenic (GM) Organisms
Direction. Match the following genetically modified organism to its
application by writing on the space given. Write the letter that corresponds to
your answer on the space before the number
 Bioethics
Bioethics is the study of controversial ethics
brought about by advances in biology and
medicine. Bioethics is multidisciplinary. It blends
philosophy, theology, history, and law with
medicine, nursing, health policy, and the medical
humanities. It is concerned with all the ethical
questions that arise in the relationships among life
sciences and the cited fields. Bioethics is
commonly understood that refers to the ethical
implications and application of the health-related
life sciences including biotechnology, such as
cloning, gene therapy, life extension, human
genetic engineering. It promotes critic reflection
about ethical conflicts, which are caused by
progressing in life science and medicine.
 Activity 4 Benefit and Risk of GMO
Direction. Write the letter B if the statement given is a benefit while the letter
R if it is a risk from the use of genetically modified organism.
________1. Crops like potato, tomato, soybean and rice are currently being
genetically engineered to obtain new strains with better
nutritional qualities and increased yield.
________2. Genetic engineering in food can be used to produce totally the
same or identical substances such as proteins and other food
nutrients.
________3. Positive genetic engineering deals with enhancing the positive
traits in an individual like increasing longevity or human capacity
while negative genetic engineering deals with the suppression of
negative traits in human beings like certain genetic diseases.
________4. Genetic engineering in food involves the alteration of genes in
crops.
________5. While increasing the immunity to diseases in plants, the
resistance genes may get transferred to the harmful pathogens.
________6. Genetic engineering can hamper the diversity in organisms.
________7. The genetic modification of foods can be used to increase their
medicinal value, thus making edible vaccines available.
 PCR Uses
• Forensics
– ID a body
– ID a criminal
– Free the innocent
• Genetic Tests
– Testing for specific polymorphism/mutation
• Paternity Tests
• Research
What is the advantage
of Covid-19 PCR-Test
vs. Rapid Test?
 PCR-Test for Covid-19
• These tests provide more accurate
results than rapid tests, and that's
because they use a polymerase chain
reaction (PCR) to identify the viral genetic
material of COVID-19.
 Gel Electrophoresis
• Gel electrophoresis is a
technique that uses gel (a thin
slab of jellylike material) as a
molecular sieve to separate
nucleic acids or proteins on
the basis of size or electrical
charge.
• This technology allows
scientists to identify
someone’s DNA!
• Watch:
• https://www.youtube.com/watc
h?v=4OJAzQsZnbo
 How gel electrophoresis would be used to separate the
various DNA molecules in three different mixtures?

– A sample of each mixture is placed in a well at one end of a flat,

rectangular gel.
– A negatively charged electrode from a power supply is attached
near the DNA-containing end of the gel, and a positive electrode is
attached near the other end.
– Because DNA molecules have negative charge owing to their
phosphate groups, they all travel through the gel toward the
positive pole.
– As they move, a thicket of polymer fibers within the gel impedes
longer molecules more than it does shorter ones, separating them
by length.
– Thus, gel electrophoresis separates a mixture of linear DNA
molecules into bands, each consisting of DNA molecules of the
same length, with shorter molecules toward the bottom.
Gel electrophoresis: a
technique used to compare
DNA from two or more
organisms.

Why compare DNA:

1. Find your baby’s daddy
2. Who committed a crime.
3. How closely species are
related.
How is
electrophoresis
done?
A. The DNA is cut into
fragments with a
restriction enzyme.
B. The cut DNA is then
put into the wells of a
machine filled with
gel.
• The gel is spongy and
the DNA squeezes
through the pores.
C. The machine is plugged in and the
fragments get separated based on their size.
• The smaller fragments move further than the
large.
 Separation of DNA based on size of
fragments.

• Electrophoresis
results

Final result of electrophoresis

• Electricity provides the energy

• Why does DNA move?

• DNA has a negative charge.

• When the machine is plugged it, its moves towards

the positive pole created by the electricity
Your DNA is so unique its considered to be a
DNA fingerprint.
Gel electrophoresis will separate your DNA
differently from anyone else.

Nova: who done it

http://www.pbs.org/wgbh/nova/sheppard/analyze.html

http://www.teachersdomain.org/asset/tdc02_i
nt_creatednafp2/
Forensic Science
 Forensic Science
• Forensic science is the scientific analysis of evidence for
crime scene and other legal investigations, and DNA
technology now plays an important role.
• In violent crimes, body fluids or small pieces of tissue may
be left at the crime scene or on the clothes of the victim or
assailant.
• If rape has occurred, semen may be recovered from the
victim’s body.
• With enough tissue or semen, forensic scientists can
determine the blood type or tissue type using older
methods that test for proteins.
• However, such tests require fresh samples in relative
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.
 Forensic Science
• DNA fingerprinting can also be used to
establish family relationships.
– A comparison of the DNA or a mother, her
child, and the purported father can
conclusively settle a question of paternity.
– Sometimes paternity is of historical interest:
DNA fingerprinting provide strong evidence
that Thomas Jefferson or one of his close
male relatives fathered at least one child with
his slave Sally Hemings.
 Forensic Science
• How reliable is DNA fingerprinting?
– In most legal cases, the probability of two people
having identical DNA fingerprints is between one
chance in 10,000 and one in a billion. The exact
figure depends on how many markers are in the
population. For this reason, DNA fingerprints are
now accepted as compelling evident by legal
experts and scientists alike.
– In fact, DNA analysis on stored forensic samples
has provided the evidence needed to solve many
“cold cases” in recent years. DNA fingerprinting
has also exonerated many wrongly convicted
people, some of whom were on death row.
Forensic Science using Gel
electrophoresis
DNA Fingerprints From a Murder Case
Genetically Modified
Organism
& Genetically
Engineered Organism
Genetically Modified Organisms
• Scientists concerned with feeding the
growing human population are using DNA
technology to make genetically modified
organisms for use in agriculture.
• A Genetically Modified Organism (GMO) is
one that has acquire one or more genes by
artificial means rather than by traditional
breeding methods. (The new gene may or
may not be from another species).
 Genetically Modified Organisms
(GMO) are called transgenic
organisms.
TRANSGENIC ANIMALS

1. Mice – used to study human

immune system
2. Chickens – more resistant to
infections
3. Cows – increase milk supply
and leaner meat
4. Goats, sheep and pigs – produce
human proteins in their milk
 Transgenic Organisms
• Organisms altered by genetic engineering.
• Genetic material changed by other than
random natural breeding
• Gene transfer
-moving a gene from one organism to another.
 What Transgenic means
• 'Trans-' means 'crossing from one place to
another‘
• The '-genic' bit means genes
• So it means that bits of genes from
different living things have been bolted
together and spliced into another organism
to make a new one which does something
which the scientists want it to do.
 Examples of Transgenic Organisms

• GMO- genetically modified organism

• GEO-genetically enhanced organism

https://www.healthproductsguru.com

For example
Plants that resists a particular type of weed killer
Sheep which makes some special substance in
its milk.
 Transgenic Goat

This goat contains

. a human
gene that codes for a blood
clotting agent. The blood
Human DNA in
a Goat Cell clotting agent can be harvested
in the goat’s milk.
 Animals used in GE
• The human gene to clot blood has been
inserted into the DNA of sheep

• Sheep produce human clotting factor

needed for Haemophiliacs in their milk

• Goats produce a protein to treat

emphysema
How to Create a Transgenic Animal?
Genetically Modified Organisms
• Transgenic animals
– The goal is, for example, to make sheep with
better quality wool or a cow that will mature in a
shorter time.
– Scientists might identify and clone a gene that
causes the development of larger muscles (which
make up most of the meat we eat) in one variety
of cattle and transfer it to other cattle or even
sheep.
– Also may be used as pharmaceutical “factories”
to produce otherwise rare biological substances
for medical use
• For example, manipulating chicken eggs.
 The DNA of plants and animals
can also be altered.
PLANTS
1. Disease-resistant
and insect-resistant
crops
2. Harder fruit
3. 70-75% of food in
supermarket is
genetically modified.
Genetically Engineered Crops
3 Types of Resistance
• Herbicide Resistance (HR)
– Most U.S. crops engineered with resistance to
glyphosate

• Insect Resistance (IR)

– Types of soil bacterium (Bacillus thuringiensis)
introduced into plant to target susceptible insects
– Ex. BT Talong
• Virus Resistance

The Impact of Genetically Engineered Crops on Farm Sustainability in the United States
Division on Earth and Life Study,The National Academes, US April 13, 2010
Facts about Genetic Engineering
• The result is called genetically modified
organism (GMO)
• One of the major aims is to produce much food
at low cost to reduce the world hunger.
• Conglomerates are buying up biotech start-up
companies, seed companies, agribusiness and
agrochemical concerns, pharmaceutical,
medical and health businesses, and food and
drink companies creating life sciences
complexes to fashion bio-industrial world
• Can potentially be used in humans to change
their appearance, intelligence, character and
adaptability???? – ethical issues
 Genetically Modified Organisms
• To make genetically modified plants, researchers can manipulate
the DNA of a single somatic cell and then grow a plant with a new
trait from the engineered cell.
– Already in commercial use are a number of crop plants
carrying new genes for desirable traits, such as delayed
ripening and resistance to spoilage and disease.
– The majority of the American soybean and cotton crops are
genetically modified.
– Many plants have received bacterial genes that make them
resistant to herbicides.
– Health benefits include “Golden rice” which produces grains
containing beta-carotene, which our body used to make
vitamin A.
• This could help prevent Vitamin A deficiency—and resulting
blindness—among the half of the world’s people who
depend on rice as their staple food.
 Genetically Modified Organisms
• Social concerns:
– Early concerns focused on the possibility that
recombinant DNA technology might create new
pathogens.
• One safety measure is a set of strict laboratory
procedures designed to protect researchers from
infection by engineered microbes and to prevent the
microbes from accidentally leaving the laboratory.
– Today, most public concern about possible
hazards centers not on recombinant microbes but
on genetically modified (GM) crops.
• Advocates of a cautious approach fear that some crops
carrying genes from other species might be hazardous to
human health or the environment.
• One specific concern is that genetic engineering could
transfer allergens to plants people eat.
 Pros & Cons of GE
Is GE precise enough ?
Pro arguments: Contra arguments:

• Scientits use „gene • The choice of gene is

guns“ to insert the precise. But the
specific gene in the insertion of this gene
organism precisely into a living cell is
imprecise. There is no
control where in the
DNA the new gene is
inserted. This process
can disrupt the DNA
 Pros & Cons of GE
Is Genetic Engineering safe?
Pro arguments Contra arguments
• All genetically engineered • Tests are only conducted on
foods have been thoroughly animals like rats and mice.
tested and demonstrated to Apart from that the scientists
be safe before they are are often not independent
due to the fact that they are
released into the involved into the big
marketplace companies

• Genetically engineered • The consequences are now

foods have been sold in the unknown and unanticipated
United States for several
years and it is no evidence • The consequences for the
to indicate that these foods human health can only be
have harmed human health assessed after human
in any way testing.
 Pros & Cons of GE
Effects on the environment
Pro arguments: Contra arguments:
• GE minimizes soil • Every genetically engineered
organism released into the
erosion by environment is a threat to the
reducing the need ecosystem because they are
of flowing . unpredictable by interacting
with other living things in the
environment, therefore it is
difficult to assess the threats of
genetically engineered
• Plants resistant to organisms to the ecosystem.
weather , climate
insect infestation, • GE can create toxins,
noxious-vegetation, harm to
desease, molds wild life and may create new
and fungi. molds and fungi.
Pros & Cons of GE
Effects on the environment
Pro arguments: Contra arguments:
• GE allows the • Once GMOs are
creation of thousands released into the
of novel life forms in a environment they
brief moment. cannot be recalled
therefore they are a
very dangerous kind
of pollution.
 Novel Products
Produced through
Genetic
Engineering
 Microorganisms
• Bacteria can make human insulin
• This prevented many diabetics from
getting an allergic reaction to animal
insulin
• Bacteria make interferon which can fight
virus infections and some cancers
 Applications (Micro-
organisms)
Production of
humulin

www.healthtap.com

Used by diabetics
74
 Plant Application
• Weedkiller resistant crops
- Weeds die but the crops survive
• Vitamin A in Rice
- The gene which produces vitamin A was
taken from daffodils and put into rice to
help prevent blindness
 Plant Application
Golden Rice – is any
variety of rice which makes
beta-carotene, thus giving
the rice a yellow (Golden)
color. It was created as an
additional intervention
for vitamin A deficiency.

76
 Bt Talong
• Bt eggplant contains a
natural protein from
the soil bacterium
Bacillus
thuringiensis which
makes it resistant to
eggplant fruit and
shoot borer (EFSB),
the most destructive
pest of eggplant
 Vaccines
• Genetically
engineered microbes
can be used to
produce the antigens
needed in a safe and
controllable way.
• The use of genetically
modified yeast cells to
produce a vaccine
against the hepatitis B
virus has been a
major success story.
 Gene Pharming
• Gene pharming is a technology that
scientists use to alter an animal's own DNA,
or to splice in new DNA, called a transgene,
from another species.
• In pharming, these genetically modified
(transgenic) animals are mostly used to
make human proteins that have medicinal
value. The protein encoded by the transgene
is secreted into the animal's milk, eggs or
blood, and then collected and purified.
 Tracy the Sheep

• One of the first mammals engineered successfully

for the purpose of pharming was a sheep named
Tracy, born in 1990 and created by scientists led
by British developmental biologist Ian Wilmut at
Roslin Institute in Scotland. Tracy was created
from a zygote genetically engineered through
DNA injection to produce milk containing large
quantities of the human enzyme alpha-1
antitrypsin, a substance used to treat cystic
fibrosis and emphysema
 Gene Therapy
• It involves modifying
human DNA either to
repair it or to replace a
faulty gene.
• The idea of gene therapy
is to overcome the
effects of a mutation
which causes a genetic
disease.
• Cystic fibrosis is the best
known disease where
gene therapy has been
tried.
 Gene Therapy
• Although the concept of gene therapy
remains promising, very little scientifically
strong evidence of effective gene therapy
has yet appeared.
• Active research into human gene therapy,
with new, tougher safety guidelines,
continues.
 Xenotransplantation
• Xenotransplantation is the
transplantation of living cells, tissues or
organs from one species to another.
• However there are ethical issues and
issues with rejection
• There are also issues with virus
transmission from one species to another
• Porcine islet transplants are being
investigated for use in type 1 diabetes due
to the shortage of human islet cells
 Diagnostic Tests
Genetic engineering can produce very
specific and sensitive diagnostic tests for
many diseases, using engineered
proteins.

This new technology is also opening up

novel ways of delivering medicines to
specific targets.