UNIT 2: MOLECULAR

BIOLOGY
Genetic engineering
Genetic engineering
Genetic engineering
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1. RESTRICTION
ENZYMES
Proteins that perform the
function of molecular scissors,
cutting specific sequences of
DNA. Each enzyme recognizes
one or a few target sequences
and cuts DNA at or near those
sequences.
Discovered in 1950s.
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1. RESTRICTION
ENZYMES
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1. RESTRICTION
ENZYMES
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2. LIGASE ENZYME
Ligase is an enzyme that can catalyze the joining of
two large molecules by forming a new chemical bond.
In genetic engineering they’re used to join DNA
molecules together to form recombinant DNA.
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2. LIGASE ENZYME
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3. VECTORS and RECOMBINANT DNA

*A vector in Genetic Engineering is a DNA molecule

used as a vehicle to artificially carry foreign genetic
material into another cell.
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3. VECTORS and RECOMBINANT DNA

*A vector in Genetic Engineering is a DNA molecule

used as a vehicle to artificially carry foreign genetic
material into another cell.
*A vector containing foreign DNA is termed
recombinant DNA. This is formed by laboratory
methods and it is possible because DNA molecules
from all organisms share the same chemical
structure.
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3. VECTORS and RECOMBINANT DNA

*A vector in Genetic Engineering is a DNA molecule

used as a vehicle to artificially carry foreign genetic
material into another cell.
*A vector containing foreign DNA is termed
recombinant DNA (recombinant DNA= foreign
DNA + vector).
This is formed by laboratory methods and it is
possible because DNA molecules from all organisms
share the same chemical structure.
The DNA sequences used in the construction of
recombinant DNA molecules can originate from any
species. For example, plant DNA may be joined to
bacterial DNA, or human DNA may be joined with
fungal DNA.
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3. VECTORS and RECOMBINANT DNA

Recombinant DNA is a piece of two stranded DNA

that has been created by combining at least two
strands which may come from different species!
Recombinant DNA and cloning
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3. VECTORS and RECOMBINANT DNA

The foreign gene is therefore called transgene (a

gene that has been transferred by genetic
engineering techniques from one organism to
another) and the organism transgenic or GMO
(genetically modified organism)
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3. VECTORS and RECOMBINANT DNA
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3. VECTORS and RECOMBINANT DNA

The most common vectors:

*Plasmid:small double-stranded and generally

circular DNA molecule that can replicate
independently.

*Viruses: genetically engineered viruses carrying

modified viral DNA or RNA that is noninfectious.

*Artificial chromosomes: can carry a much larger

DNA fragment than other vectors
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3. VECTORS and RECOMBINANT DNA
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3. VECTORS and RECOMBINANT DNA
https://www.yourgenome.org/facts/what-is-genetic-e
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4. PCR: polymerase chain reaction

PCR is a method widely used in molecular biology in

order to obtain several copies of a specific DNA
segment. Using PCR, copies of DNA sequences are
exponentially amplified to generate thousands to
millions of more copies of that particular DNA
segment. PCR is now a common and often
indispensable technique used in medical and clinical
laboratory research for a broad variety of applications
including biomedical research and criminal forensics.
Developed in the 1980s.
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4. PCR: polymerase chain reaction
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4. PCR: polymerase chain reaction
Polymerase chain reaction (PCR)
Kary Mullis (1983)
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STEM CELLS: células troncales o células madre

TOTIPOTENTES: a.k.a. omnipotentes. Estas células madre

pueden diferenciarse en tipos celulares embrionarios y
extraembrionarios. Son capaces de construir un organismo
completo y viable. Se originan tras la fusión de un óvulo y un
espermatozoide (cigoto). Esta totipotencia se conserva tras
las primeras divisiones del óvulo fertilizado (o cigoto)
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STEM CELLS: células troncales o células madre

PLURIPOTENTES estas células troncales son las descendientes

de las totipotentes y pueden diferenciarse en prácticamente
todas las células, pero no pueden dar un organismo entero.

MULTIPOTENTES pueden diferenciarse en varios tipos

celulares, pero pertenecientes a la misma “familia” de células.
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STEM CELLS: células troncales o células madre

OLIGOPOTENTES estas células sólo pueden diferenciarse en unos

pocos tipos celulares, como las células madre linfoides o mieloides
del tejido óseo.

UNIPOTENTES estas células sólo pueden originar un tipo celular,

siendo capaces de autorrenovarse.
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STEM CELLS
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STEM CELLS
 ANATOMÍA APLICADA

Tejido Tipos
 ANATOMÍA APLICADA

Tejido Tipos
Tejido
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5. CLONING

Cloning is obtaining identical copies of a gene or an

entire organism. It’s useful, for example, for
producing large amounts of proteins or human insulin
using the human gene (transgene) inserted in a
plasmid (vector) inside bacteria (GMO).
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5. CLONING
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5. CLONING: USES
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5. CLONING
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5. CLONING: USES

Therapeutic cloning

Reproductive cloning
Animal cloning

Therapeutic cloning Reproductive cloning

Therapeutic cloning

Stem or mother
cells
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5. CLONING: USES

Therapeutic cloning
Cloning tissues or organs in order to provide ill patients with
a transplant.
The nucleus of a cell from the patient is introduced into an
ovum/oocyte (from a different donor). The result is an
embryo with the same genes of the patient.
The oocyte with the nuclear transfer develops into a
blastocyst (blastula) made of stem cells that shall be used
to obtain the tissue or organ that we want to transplant,
100% compatible with the patient (donor and recipient are
the same person!)
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5. CLONING: USES
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5. CLONING: USES

Reproductive cloning
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5. CLONING: USES

Reproductive cloning
Involves cloning individuals that are genetically identical.
Risks: individuals age faster and develop illnesses. Prohibited in humans.

1. Immature egg (oocyte) with nucleus removed (Mother 1)

2. Nucleus of oocyte replaced by nucleus of a somatic cell (Mother 2)
3. Embryo (oocyte with the replaced nucleus) implanted in Mother 3
which goes through pregnancy.
4. The sheep born is identical to the nucleus donor (Mother 2).
Reproductive cloning

Dolly the sheep

1996-2003
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5. CLONING: USES
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5. CLONING: USES
https://www.youtube.com/watch?v=qfNUGN
Genetically modified organisms (GMO)
Plants that are insect-resistant
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6. DNA HYBRIDIZATION
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6. DNA HYBRIDIZATION
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6. DNA HYBRIDIZATION
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6. DNA HYBRIDIZATION
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6. DNA HYBRIDIZATION
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6. DNA HYBRIDIZATION
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7. GENE DELIVERY: WAYS TO DELIVER TRANSGENES

• Bacterial transformation
• Heat shock
• Liposome
• Nanoparticles
• Electroporation/Sonoporation/Photoporation/Magnetofection
(electricity/sound waves/laser/magnetic fields)
• Biolistics (gene gun)
• Microinjection
• Agrobacterium (bacteria) delivery
• Viral delivery
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7. GENE DELIVERY: WAYS TO DELIVER TRANSGENES

Bacterial transformation
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7. GENE DELIVERY: WAYS TO DELIVER TRANSGENES

Agrobacterium
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7. GENE DELIVERY: WAYS TO DELIVER TRANSGENES

Agrobacterium
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7. GENE DELIVERY: WAYS TO DELIVER TRANSGENES

Viral delivery
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7. GENE DELIVERY: WAYS TO DELIVER TRANSGENES

Biolistics: Gene gun

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7. GENE DELIVERY: WAYS TO DELIVER TRANSGENES

Biolistics: Gene gun

Evolución de los cultivos transgénicos en hectáreas (Unión
Europea)
https://es.greenpeace.org/es/trabajamos-en/agricultura/transg
enicos/
https://es.greenpeace.org/es/trabajamos-en/agricultura
https://www.telesurtv.net/news/Los-12-productos-mas-nocivos-de-M
onsanto-Fotos--20141022-0037.html

https://es.wikipedia.org/wiki/Monsanto
https://www.lavanguardia.com/comer/materia-prima/2
0180424/442902035279/transgenicos-pros-contras.ht
ml
 GMO/ TRANSGENICS

Use of GMOs:

• Bacteria for human insulin production

• Viruses with human genes for gene therapy
• Modified animals to enhance their traits: e.g milk
production
• Edible plants resistant to herbicides, plagues or with
higher vitamin content.
 GMO/ TRANSGENICS

Use of GMOs:

• Bacteria for human insulin production

• Viruses with human genes for gene therapy
• Modified animals to enhance their traits: e.g milk
production
• Edible plants resistant to herbicides, plagues or with
higher vitamin content.
• IT’S NOT POSSIBLE TO BRING DINOSAURS BACK TO
LIFE 
 GMO/ TRANSGENICS

Use of GMOs:

• Bacteria for human insulin production

• Viruses with human genes for gene therapy
• Modified animals to enhance their traits: e.g milk
production
• Edible plants resistant to herbicides, plagues or with
higher vitamin content.
• IT’S NOT POSSIBLE TO BRING DINOSAURS BACK TO
LIFE 
 GMO/ TRANSGENICS

Problems with GMOs:

• Environmental issues: wild type varieties extinction.

• Could increase inequality between rich and poor countries
(agriculture and economies).
• Monopoly of biotech corporations.
• Not scientifically proved that may cause cancer.
GMO/ TRANSGENICS
 Biotechnology

It’s the discipline that employs technology in biological procedures. It

uses living organisms in order to obtain particular products or
processes.
 Biotechnology

Traditional biotechnology uses microorganisms for completing or

increasing the efficiency of production processes.
 Biotechnology

There are three main areas in which traditional biotechnology is

used: food, healthcare and the environment.
 Biotechnology

Food:

• Bread and beer, products of fermentation of cereals by the yeast

(microscopic fungus) Saccharomyces cerevisiae.
• Cheese and dairy products (yoghourt) made by fermentation by
bacteria species such as Lactobacillus, Lactococcus or
Streptococcus or moulds such as Penicillium (blue cheese or
Roquefort)
 Biotechnology

Healthcare:

• Antibiotics (substances which fight bacteria) produced by

moulds such as Penicillium. Penicilin was the first antibiotic ever
to be discovered. Alexander Fleming won the Nobel Prize for that
Biotechnology
 Biotechnology

Healthcare:

• Vaccines are substances used as a preventive measure against

viral and bacterial infections and are made by inactive/weakened
microorganisms or fragments of them.
Biotechnology
Biotechnology

Herd immunity
Biotechnology

Vaccines
Biotechnology

Vaccines
Biotechnology

Vaccines
Biotechnology

Vaccines
 Biotechnology

Environmental:

• Bacteria for decomposition of urban waste or sewage by

fermentation (in EDAR)
• Bioplastics: biodegradable materials.
• Elimination of oil spills by bacteria (sometimes GMO).
 Biotechnology

Modern biotechnology is based on the achievements of genetic

engineering. It started in the 1980s (RE, PCR…) and it is used
along with traditional biotechnology to make food, medicines,
agricultural and industrial products.
 Biotechnology

Farming: improvement of food production by GMOs resistant to

infestations or herbicides or the increase of beneficial traits (e.g:
vitamin content).
This is achieved by cloning and transgenic techniques (previously
explained).
 Biotechnology

Biohealth:

1. Molecular diagnosis (markers).

2. New medicines and vaccines (low adverse side-effects).
3. Cell and tissue engineering (stem cells).
4. Gene therapy: flawed proteins replacement.
 Biotechnology

Environmental:

• Bacteria for decomposition of urban waste or sewage by

fermentation (in EDAR)
• Bioplastics: biodegradable materials.
• Elimination of oil spills by GMO bacteria
 BIOETHICS

IMPLICATIONS:

• Ethical: biopatents (“playing God”)

• Social (differences rich vs poor)
• Environmental (disappearance of wild types)

Bioethics is a discipline created in 1977 to combine technology,

knowledge and values. The bioethical principles are:

Justice
Non maleficence
Beneficence
Autonomy
BIOETHICS