Syllabus Reference:

5.8.1 9
Discuss how the outcomes of the Human Genome Project are being used in the development of new drugs & the social, moral and ethical issues this raises.

The Human Genome Project (identifying the complete sequence of code in human DNA) has initiated a new field of medicine called pharmacogenetics - the idea that drugs can be targeted at / tailored for individuals based on their DNA. This would allow: 1) Correct / lower dosages to be used (costs less and fewer sideeffects - e.g. cytochrome P450 gene) 2) Ability to choose drugs that work best for individuals (e.g. common painkillers are more effective for pale skinned, red-haired woman; ibuprofen does not work as well during 2nd half of menstrual cycle) 3) Ability to avoid drugs that wont work in individuals - e.g. kappaopioids (pain killers) work better on women than men. 4) Ability to avoid drugs that would give adverse reactions (would save lives and money)

Pharmacogenomics example now in use: Cytochrome P450 gene This is a gene that codes for liver enzymes which break down drugs so that they can be eliminated from the body. Gene variations mean some individuals have less P450 and are so are vulnerable to drug overdoses. Genetic tests can check the P450 gene variation an individual has, so that the correct dosage can be given.

There is a need to consider the social, moral & ethical issues of HGP & pharmacogenomics:
Pharmacogenomics: 1) Identifying the correct genes is time consuming and 5.8.1 costly- is the potential benefit worth the cost? 1 2) Is it ethical to leave people with certain genetic variations without a particular treatment or drug? 3) Is it worth the financial implications in further training for doctors? 4) DNA databases will need to be analysed and stored - is it ethical to use these for criminal investigations / immigration issues? Human Genome Project:
5) Can you patent / copyright DNA when you sequence it? 6) Who should have access to your genetic code? 7) Should genetic information be kept on record? 8) What if we identify genes for intelligence, sexuality, crime? 9) Eugenics (i.e. selective breeding in humans) - is this ethical?

Syllabus Reference:
5.8.2 0
Describe how drugs can be produced using GM organisms (plants, animals & micro-organisms).

Maize Has been modified to protect against the corn borer using bacteria (see next slide)

GM Plant crops
Soya Bean Plants Have been made herbicide resistant, so they can be sprayed without being affected.

Flavr Savr Tomatoes have been modified to slow down the ripening process, using an 'antisense' gene (which blocks enzyme gene expression).

Oilseed Rape has been GM to produce a different oil, using a bacterium called agrobacterium. (see next slides)

Simplified!!

Genetic Engineering: How to make a transgenic organism (i.e. with DNA from more than one source)
5.8.1 1

More Detailed!!

Risks of Genetic Engineering

- Can have side effects (see below)! - It's very difficult (so LOTS of repeats to get one working attempt - bad if we're engineering animals) - Costly - In crops the genes often 'escape' into other plants which may lead to immunity and infertility. - Opens door to human GE - Accidental release of GE organisms into the wild is BAD - GE biological weapons - Religious objection

Electrophoresis

A form of chromatography used to separate

different pieces of DNA on the basis of their length

Making human Insulin - using transgenic micro-organisms Type 1 & 2 diabetes sufferers need regular insulin injections Used to come from animals such as pigs and cows Now micro-organisms can be used to manufacture insulin (aka 'humulin'). This can be cultured in huge numbers, though downstream processing is used to separate the end product from the mixture.

Human Insulin production - how it works:

GM Microbes - Banana Vaccines

Plants (such as bananas or carrots) can be GM to carry vaccines to human diseases, e.g. diarrhoea or hepatitis B. Plants can be regrown by communities, cheaply and easily. Plants do not need a cold storage No need for trained Healthcare workers No need for needle injections.

Making Transgenic Animals Getting the DNA in!

As well as using a plasmid, there are other ways of introducing new DNA into a eukaryotic cell: Different vectors may be used: 1) Harmless viruses - used to infect the animal cell.
2) Liposome wrapping - inserted gene is wrapped in a liposome which fuses with the cell membrane to deliver the DNA. 3) Microinjections - DNA is injected into the cell using a fine micropippette. Used to produce 'Tracey' - the transgenic sheep.

4) Microprojections - using a gene gun - DNA is attache to tiny gold pellets and fired into the cell using compressed air.

Making Transgenic Animals Microprojections

1) Coat gold particles with DNA vectors.

2) Load onto a pellet

3) Load pellet into a gene gun

4) Shooting the gene gun releases the particles at a high velocity, penetrating the cells.

AAT High yields from transgenic sheep. Help to treat genetically linked emphysema & CF

Activated protein C Used for treating DVT

Drugs from transgenic animals

Activated protein C Produces modified milk for lactose intolorence sufferers

Blood Factor VII & IX Important for blood clotting. Help haemophiliacs. Harvested from transgenic milk

Tracey - the Transgenic Sheep

1991 - Tracey the 1st transgenic sheep was born, after the gene alpha-1-antitrypsin (AAT) had been inserted into her DNA. Tracey produced the human protein AAT in her milk. Tracey's milk can be harvested & purified to extract AAT & given to people without the AAT protein (suffering from lung & liver conditions causing emphysema).

'Pharming' Facts
Using transgenic animals is sometimes called 'Pharming' Such methods gave rise to Dolly the sheep (the first clone) Research is under way to grow human organs (such as kidneys) using living pigs. The animals used are like 'living factories' for human proteins or organs.

www.mnn.com/green-tech/research-innovations/photos/12bizarre-examples-of-genetic-engineering

Syllabus Reference:
Discuss the risks and benefits 5.8.21 associated with the use of GM organisms.

How to make a transgenic organism (i.e. with DNA from more than one source) 1. Cut required gene out of donor cell DNA (using a restriction enzyme). 2. Separate gene from other fragments of DNA using electrophoresis & a gene probe complementary to the gene 3. Extract the gene from the gel and copy it using PCR (Polymerase Chain Reaction - allows many copies of DNA to be made from a small sample) 4. Extract plasmids from bacteria (macerate & centrifuge) 5. Cut plasmids open using the same restriction enzyme.

6. Insert the gene into plasmid and seal with DNA Ligase enzyme
7. Use a vector to get the transgenic plasmid back into a bacterium (vector is usually heat shocked)

Benefits of Genetic Engineering - Can manufacture desired protein easily (e.g. Insulin) - Most proteins can't be made synthetically - Treatment of diseases - Low running costs (after initial cost of making GE organism). - Renewable - New technology available (e.g. body armour made from spiders silk produced by GE goats)