Biotechnology CLIL module

Biotechnology

CLIL MODULE ABOUT…

Author: Prof.ssa Elena Bruno

UNIT 1CONTENTS

• What’s biotechnology?• How old is biotechnology?• Differences between

traditional and innovative biotechnology

• Recombinant DNA technology

• Production of insulinwith recombinant DNA technology

What is biotechnology?

Biotechnology is the use of living cells or organisms to produce materials useful to people.

TraditionalBiotechnology

Innovative Biotechnology

Use of living beings and selection ofnatural features in living beings

use of genetically engineeredmicroorganisms, plant and animal cells

GMO

How Old Is Biotechnology?10,000 BCDomesticating Crops

1880’s Production of Vaccines

Domesticating Animals8,000-9,000 BC

4,000 BC Leavening Bread

6,000 BC Brewing Beer

1940’s Production of Antibiotics

1980’s Use of genetically modified organisms

molds

yeasts

Do traditional biotechnologymodify living beings?

Old biotechnology modifies living beings by selective breeding ofplants/animals: on the contrary of modern biotechnology, scientists do notdirectly modify the DNA

Do traditional biotechnologymodify living beings?

The traditional biotechnology also use radiations (such as X-rays) to increasethe rate of mutations in the DNA of a species (such as wheat): in this way it ispossible to increase the genetic variability in a short time and to obtainfavorable variations. For example the variety "wheat Creso" was obtained inthis way.

irradiated wheat is not a GMO!

Do innovative biotechnologymodifies living beings?

Modern biotechnology modifygenetic material (DNA) in a waythat does not occur naturally bymating and/or naturalrecombination

EXAMPLE: Resistance against insects is achieved by incorporating into the food plant the gene for toxin production from the bacterium Bacillus thuringiensis (Bt)

CORN COB

Applications of biotechnologyMedical Biotechnology• Diagnostics• Therapeutics• Vaccines

AgriculturalBiotechnology• Plant agriculture• farms• Food processing

EnvironmentalBiotechnology• Cleaning through

bioremediation• Preventing

environmentalproblems

Forensic Biotechnology• Paternity test• Scientific

investigations

Industrial Biotechnology• Energy production• New materials

APPLICATIONS OF BIOTECHNOLOGY

Some tools of biotechnologyRecombinant DNA technologyGene TherapyCloningMonoclonal Antibodies’productionStem cellsPCRDNA fingerprintDNA sequencingIn vitro fertilization……AND MORE!!!!

Recombinant DNA technologyRecombinant DNA technology is one of the recent advances in

biotechnology, which was developed by two scientists named

Boyer and Cohen in 1973.

Recombinant DNA technology works by taking DNA from two

different sources and combining that DNA into a single molecule.

That alone, however, will not do much. Recombinant DNA

technology only becomes useful when that artificially-created

DNA is reproduced. This is known as DNA cloning.

What’s recombinant DNA?RECOMBINANT DNA : is a DNA molecule that has been made

in the laboratory using at least two different sources of DNA

Recombinant DNA is produced using in vitro molecular

techniques to isolate and manipulate fragments of DNA

?

To produce Recombinat DNA you need

Molecular scissors: Restriction enzyme

Molecular glue: Ligasi enzyme

A vector:Plasmide, viruses,

etc

Restriction enzymesEnzymes that cleave DNA are used to manipulate and recombine

DNA.

Restriction enzymes, naturally used by bacteria as defense against

bacteriophages, cut DNA into smaller pieces.

Restriction enzymes cut DNA at

specific base sequences called

restriction site.

CUT=CLEAVE

Restriction enzymesSome restriction

enzymes

recognise and

cut palindromic

DNA sequences

(sequences that

read the same

way in both

directions).

Some of them

make staggered

cuts, generating

fragments with

sticky ends

Restriction enzymes

Sticky ends

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Enzyme ligaseThe sticky ends may join: a fragment from one source, such as human, canbe joined to a fragment from another source, such as bacterium. Initiallythe fragments are held together only by weak hydrogen bonds: for thesereason you use the enzyme ligase to catalyze the formation of covalentbonds between adjacent nucleotides at the ends of the fragments, joiningthem to form a single, larger molecule

They join

ligase

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Enzyme ligase

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PlasmidsA plasmid is a small DNA molecule within a cell that is physically separatedfrom a chromosomal DNA and can replicate independently. They are mostcommonly found in bacteria as small, circular, DNA molecules• Plasmids are naturally exchanged among organisms• They can be modified and used to deliver new pieces of DNA into anorganism=VECTORThey often contain genes for antibiotic resistance

http://education-portal.com/academy/lesson/what-is-a-dna-plasmid-importance-to-genetic-engineering.html#lesson

Six steps of recombinant DNA technology

1. Isolating (vector andtarget gene)

2. Cutting (Cleavage)

3. Joining (Ligation)

6. Selecting (Screening)

5. Cloning

2

3

1

4

4. Transformation(Introduction of the plasmid intocells) 5

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Six steps of recombinant DNA technology

Screening

ampicilline resistance gene

lacZ gene

unfunctioning lacZ gene

human gene

Plate out the transformedbacteria on a solid nutrientmedium containing ampicillinand a sugar called X-gal.

Only transformedbacteria that havethe ampicillin-resistance plasmidwill grow.

This bacterium express lacZgene, willhidrolyse sugar and form BLUE colony

This bacterium DOESN’T express lacZgene, it isn’t able to hidrolyse sugar, will form WHITE colony: it’s a GMO with useful external gene

Repotergenes or marker genes

Screening

Production of insulin in the pastType I diabetes mellitus is a metabolic diseasecaused by lack of hormone insulin.It’s treatable only with injection

In the past, insulin was taken from the pancreasesof cows and pigs. The method was expensive andsome patients had severe allergic reaction to theforeign insulin or its contaminants

these problemshave beenovercome thanksto a GMO

http://education-portal.com/academy/lesson/what-is-genetic-engineering-definition-and-examples.html

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Production of insulin with recombinat DNA technology

simplifiedversion

Production of insulin with recombinat DNA technology

Correctversion

Two genes are chenically synthesized, onefor the insulin A chain and one for theInsulin B chain.Plasmid are extracted from Escherichiacoli. To make the bacteria produce insulin,the insulin gene must be linked to thebeta-galactosidase gene, which carries apromoter for tanscription.The recombinant plasmids are insertedback into bacteria, these bacteriareproduce so that Insulin gene are cloned.The GMO bacteria produce the two Insulinchains that are extracted, purufied andmixed together .The A and B insulin chains conncet viadisulfade cross linkages to form thefunctional insulin protein

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Animals are genetically engineered to exalt specific traits or

obtain useful products:

•GM animals can produce pharmaceuticals

•GM mice are bred for human disease research

•Xenotransplantation (animals as organs donor in

transplantation)

Genetically modified animals

Genetically modified animals

Genetically modified animals

UNIT 2CONTENTS

• Biotecchnology in medicine: production of drugs

• Monoclonal antibodies• Stem cells (blended

learning)• Gene therapy

TPA is a naturally occuring human protein

that dissolves blood clots. It’used to treatpatients suffering from blood clotting in heartattack or strokes

Production of drugs

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Production of drugs

Production of drugs with pharming

An expression vector

carrying a desired gene can

be put into an animal

fertilized egg (embryo),

which is implanted into a

surrogate mother. The

transgenic offspring

produce the new protein in

their milk. The milk is easily

harvested and the protein

isolated, purified, and made

clinically available to

patients.

Another way of making medically useful products in large amount is

pharming: the production of pharmaceuticals in farm animals or plants

2. The human transgene is injectedinto the Fertilized eggs

3. Eggs are transferred to recipient ewes

1. Donor ewes: afterinsemination, fertilized eggs are collected

4. The offspringare raised-selection of animalsproducing the human protein in the milk

Ewe=female sheep

Production of drugs with pharming

• Human growth hormone (forchildren suffering deficiencies) cannow be produced by transgeniccows.

• Only 15 such cows are needed tosupply all the children in the worldsuffering from this type ofdwarfism.

transgenic animals are not well seen by the public, and yet ....

Monoclonal antibodies

https://www.youtube.com/watch?v=Qp0z3bZDfKE

Use of Monoclonal antibodies

Use of Monoclonal antibodies

ADC: monoclonal antibodies drug conjugated

https://www.youtube.com/watch?v=ACxmsMmo9hU

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Production of vaccinesTypes of Vaccine

Whole-Agent Vaccine: Contains whole, non virulent microorganisms

Subunit vaccine: Contains some partor product of microrganisms that canproduce an immune response

ProblemsThese microrganisms can back-mutate to a virulent formAllergic reactions (e.g. against eggsubstrate used to grow viruses)

They can be produced withbiotechnology, they are safer thanattenuated vaccines because theycan’t reproduce and they containlittle or no extra material (no allergicreactions)

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Production of recombinant vaccines

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Production of vaccines

Gene TherapyGene therapy is an experimental technique for treating disease by altering the patient'sgenetic material. Most often, gene therapy works by introducing a healthy copy of adefective gene into the patient's cells.

Researchers must find safe, efficient vehicles--called vectors--to carry the modified DNA to targeted cells. There are several vectors already in use, including chemical solutions, synthetic fat molecules, and viruses that have been modified so that they are harmless.

Gene TherapyGene therapy may be classified into the two following types, only one of which has been used in humans:

Somatic gene therapyAs the name suggests, in somatic genetherapy, the therapeutic genes aretransferred into the somatic cells (non sex-cells), or body, of a patient. Anymodifications and effects will be restrictedto the individual patient only, and will notbe inherited by the patient's offspring orlater generations

Germline gene therapyIn germline gene therapy, germcells (sperm or eggs) are modified by theintroduction of functional genes, whichare integrated into their genomes. Germcells will combine to form a zygote whichwill divide to produce all the other cellsin an organism and therefore if a germcell is genetically modified then all thecells in the organism will contain themodified gene. This would allow thetherapy to be heritable and passed on tolater generations

technical and ethicalproblems=eugenics

Gene Therapy

Gene Therapy to treat SCID

A form of SCID is caused by a deficiency of an enzyme (adenosine deaminase or ADA), which is necessary for lymphocytes to develop.

http://learn.genetics.utah.edu/content/genetherapy/gtdoctor/

Space DoctorWelcome to the Extraterrestrial Gene Therapy Lab! As Space Doctor, you'll use your gene therapy knowledge to treat three ailing aliens who come into your clinic.

A bit of fun…with gene therapy

Stem cells

https://www.youtube.com/watch?v=2-3J6JGN-_Y

ESL VIDEO: A Stem Cell Story Quick Quiz

UNIT 3• Pro and cons of biotechnology

PRO: advantages of GMOsInsect ResistanceSome GMO foods have been modified to make them more resistant to insect pests.The University of California in San Diego reports that a toxic bacterium can be addedto crops to make them insect repellent, yet safe for human use. This can reduce theamount of pesticide chemicals used on the plants, thus potentially reducingexposure to pesticides.

PRO: advantages of GMOs

More Nutritious FoodsThe Food and Agricultural Organization of the United Nations reports that someGMO foods have been engineered to be more nutritious in terms of mineral orvitamin content. Not only does this help you get the nutrients you need, it can alsoplay a significant role in battling malnutrition in the developing world. The UnitedNations advises that vitamin A-enhanced rice is helping to reduce global vitamin Adeficiencies.

Golden rice

PRO: advantages of GMOs

More Nutritious Foods

Genetically-modified purple tomatoes : Their dark pigment is intended to givetomatoes the same potential health benefits as fruit such as blueberries.

PRO: advantages of GMOs

More resistant plant•Cold tolerance An antifreeze

gene from cold water fish has beenintroduced into plants such as tobaccoand potato. With this antifreeze gene,these plants are able to tolerate coldtemperatures that normally would killunmodified seedlings.

•Drought tolerance/salinitytolerance Creating plants that can

withstand long periods of drought orhigh salt content in soil andgroundwater will help people to growcrops in formerly inhospitable places

tomatoes long conservation

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Edible vaccinesPlants enginereed to contain the vaccine can be grown locally, in the area wherevaccination in required, overcoming the logistic and economic problems oftransporting prepared vaccines over long distances. Most immportantly, ediblevaccines do not require syringes, saving money and eliminating the risk ofinfection from contamined needles.

PRO: advantages of GMOs

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BioremediationPRO: advantages of GMOs

GM bacteria can clean up oil contamination repidly and effectly, but can youthink about the effect caused by the uncontrolled spread of these GM bacteria?

consumers are the guinea pigs?

CONS: problems with GMOs

Allergic ReactionsGMO foods can present significant allergy risks to people. Genetic modification oftenmixes or adds proteins that weren't indigenous to the original plant or animal, causingnew allergic reactions in the human body. In some cases, proteins from an organism thatyou're allergic to may be added to an organism that you weren't originally allergic to,prompting the same allergic reaction experienced from the first organism.

CONS: problems with GMOs

Possible «escape» of transgene from crops to other species

Gene TransferA constant risk of GMO foods is that the modified genes of the organisms may escape into the wild. Brown University warns that herbicide-resistant genes from commercial crops may cross into the wild weed population, thus creating "superweeds" that are impossible to kill with herbicides

More than tenresistant weedspecies have

appeared in the U.S

CONS: problems with GMOs

Gene TransferA related risk is that the escape of genetically enhanced animals and vegetation cancreate new super-organisms that can out-compete natural animal and plantpopulations to drive certain species into extinction.

CONS: problems with GMOs

some crop are genetically modified to resist herbicides, this encourages extensive use of herbicides. Another negative impact is the possibility that increased us of an herbicide will select for weeds with naturally mutations that make them resistant to that herbicide (such as glycofosfate)

CONS: problems with GMOs

Economic concernsBringing a GM food to market is a lengthy and costly process, and of course agri-biotechcompanies wish to ensure a profitable return on their investment. Many new plant genetic

engineering technologies and GM plants have been patented, and patent infringement is abig concern of agribusiness. Yet consumer advocates are worried that patenting these newplant varieties will raise the price of seeds so high that small farmers and third worldcountries will not be able to afford seeds for GM crops, thus widening the gap between thewealthy and the poor

CONS: problems with GMOs

CONS: problems with GMOs

Ethical issues

See-Through Frog.

GMO Bulls

GM animals may suffer poor health and reduced life span

Bald Chickens