1. What About the Term Genetic Engineering?
Genetic engineering is the basic tool set of biotechnology
Genetic engineering involves:
Isolating genes
Modifying genes so they function better
Preparing genes to be inserted into a new species
Developing transgenes
Genetic engineering is the collection of techniques necessary to create a transgene. These procedures include isolating the gene-of-interest from the tens of thousands of genes found in the genome of a species. Once that gene is isolated, it is modified so it functions better in an organism. That gene is then mixed with other genes to prepare it to be introduced into another organism. This whole step develops transgenes.

2. What is a transgenic? Concept Based on the Term Transgene
Transgene – the genetically engineered gene added to a species
Ex. – modified EPSP synthase gene (encodes a protein that functions even when plant is treated with Roundup)
Transgenic – an organism containing a transgene introduced by technological (not breeding) methods
Here I am pointing out the difference between the process (using a transgene) and the product (the transgenic plant).
Ex. – Roundup Ready Crops

3. Why are transgenics important?
We can develop organisms that express a “novel” trait not normally found in the species
Extended shelf-life tomato (Flavr-Savr)
It is important to separate the process from the product. Focus on the product, it will highlight the benefits of biotechnology.
Herbicide resistant soybean (Roundup Ready)

4. Environmental Cleanup
Biotechnology is Not Just on the Farm
Disease Treatment
Diagnostics
Environmental Cleanup
Human Applications
A list of human and environmental applications of biotechnology.

5. Human Applications Pharmaceutical products
New solutions to old problems
Disease diagnosis
Determine what disease you have or may get 
Gene therapy
Correcting disease by introducing a corrective gene
These are the general classes of human applications. Pharmaceutical products will be discussed on the next slide. Disease diagnosis using the tools of biotechnology to determine if an individual is at risk for contracting a certain disease. In some cases, the result will predict a 100% probability, whereas in other cases a positive result means a significantly increased risk. Gene therapy involves the insertion of a functioning gene into a human. The gene provides a protein product that is not being produced in the individual because of a mutation in their corresponding gene.

6. Biotechnology and Health
Product
Use
Insulin
Diabetes
Interferon
Cancer
Interleukin
Human growth hormone
Dwarfism
Neuroactive proteins
Pain
Each of these products were developed by the pharmaceutical industry using biotechnology approaches. Most of these are engineered into bacteria, and the protein product is extract in large quantities for future sale.
The genes for these proteins are:
Cloned
Inserted into bacteria
Product isolated using biofermentation

7. Environmental Applications
Bioremediation - cleanup contaminated
sites; uses microbes designed to degrade
the pollution
Indicator bacteria – contamination can be
detected in the environment
Recombinant (genetically-engineered) bacteria have been developed that help clean up contaminated sites. They can also be used to determine if a specific site is contaminated with a specific pollutant.

8. Future Health-related Biotech Products
Vaccines – herpes, hepatitis C, AIDS, malaria
Tooth decay – engineered Streptococcus mutans,
the bacteria that destroys enamel
The health and pharmaceutical industry uses biotech approaches for vaccine production. The normal bacteria in the mouth produces an acid that destroys enamel. The engineered version of the bacteria does not produce the acid. Children will be treated with the engineered version, which also has a gene that will destroy the other bacteria in the mouth, allowing it to become established.

9. Transgenic Plants Serving Human Health Needs
Edible Vaccines
Transgenic Plants Serving Human Health Needs
Works like any vaccine
A transgenic plant with a pathogen protein gene is developed
Potato, banana, and tomato are targets
Humans eat the plant
The body produces antibodies against pathogen protein
Humans are “immunized” against the pathogen
Examples:
Diarrhea
Hepatitis B
Measles
Edible vaccines may be the most important and accepted biotech product. The principles are the same as those used for normal vaccines: a protein enters the body in some manner, and the human immune system produces antibodies against that protein. When the human is then exposed to the pathogen, the immune system is turned on and destroys the pathogen.

10. A Popular Term We Need To Know
GMOs - Genetically modified organisms
GMO - an organism that expresses traits that result
from the introduction of foreign DNA
Originally a term equivalent to transgenic organism
GMO: notice that foreign DNA is a requirement for this definition. Plant breeding products are not GMOs, as some would like to claim.

11. Delivering the Gene to the Plant
Transformation cassettes are developed in the lab
They are then introduced into a plant
Two major delivery methods
Agrobacterium
Two techniques are used to deliver DNA found in the transformation cassette into plant tissues during the plant transformation process. One is a biological system based on the plant pathogen Agrobacterium tumefaciens. The second is a mechanical method where the DNA is “shot” into plant cells using a gene gun.
Regardless of the delivery method, the delivery system must use a plant tissue source that can be manipulated to produce new plants.
Tissue culture
required to generate
transgenic plants
Gene Gun

12. Microinjection into the germ line -> transgenic animal
Gene injected into the male pronuclei

13. Recombinant Defective Retrovirus
Eggs are infected prior to fertilization
Virus integrates into one of the chromosomes

14. PROTOPLAST preparation

15. Gene Gun

16. The Lab Steps
And this slide illustrates those steps.

17. The Next Test Is The Field
Herbicide Resistance
Non-transgenics
The last step in plant genetic engineering is field testing. This slide shows a field that contains herbicide resistant and tolerant plants.
Transgenics

18. Final Test of the Transgenic
Consumer Acceptance
RoundUp Ready Corn
What is needed is for the public to accept these crops. Examples such as these, were a corn crop is freed of weed pressure make a compelling case for acceptance of these new agricultural products. But, it should be noted that these traits are all producer orientated.
Before
After

19. Examples of Transgenic Animals

20. Transgenic Cattle
Dairy cows carrying extra copies of two types of casein genes produce 13% more milk protein
Not only will this make the milk more nutritious, it would allow for less milk to make more cheese
Currently the milk from these animals is under FDA review
The important difference between this & other transgenics is that the DNA added is not foreign
                                                        

21. EnviroPig TM Transgenic pigs express phytase in their salivary glands
Phytic acid in the pig meal is degraded releasing phosphorus
The phosphorus is absorbed by the pig
Normally the phytic acid/phosphorus complex passes through the pig and is excreted as waste
Pig waste is a major pollutant & can cause eutrophication of lakes & streams

22. Transgenic Fish Tilapia Salmon/trout Catfish
Can grow up to 6 times faster than wildtype fish
Most have extra copies of growth hormone (GH) gene
Transgenic
Wildtype

23. Antifreeze Proteins
AFPs lower the freezing temperature of blood & fluids
Trout normally do not survive in water below –0.6°C
Transgenic trout containing an AFP gene & promoter can survive in waters as cold as –1.2°C
wild
transgenic
+Antifreeze

24. Animal Bioreactors
“Pharming”

25. alpha-1-antitrypsin (AAT) treatment for emphysema & cystic fibrosis
1997, Tracy the sheep, the first transgenic animal to produce a recombinant protein drug in her milk
alpha-1-antitrypsin (AAT) treatment for emphysema & cystic fibrosis
Created by PPL Therapeutics & The Roslin Institute

26. The resulting male goats were used to sire silk-producing female goats
Nexia Biotechnologies transfered the silk gene from Orb spiders into goats
The resulting male goats were used to sire silk-producing female goats
Each goat produces several grams of silk protein in her milk
The silk is extracted, dried to a white powder, and spun into fibers
The fibers are stronger and more flexible than steel
Webster and Peter
Transgenic male kids carrying silk gene

27. There is interest in using rabbits since housing costs are significantly less & generation time is faster
Chickens which produce recombinant drugs in their eggs have been produced by The Roslin Institute

28. Other Types of Transgenic Animals

29. Transgene -> Gene coding for a growth hormone

30. ANDi, the first transgenic primate born in January, 2000
224 unfertilized rhesus eggs were infected with a GFP virus
~Half of the fertilized eggs grew and divided
40 were implanted into twenty surrogate mothers
five males were born,two were stillborn
ANDi was the only live monkey carrying the GFP gene
                                                

31. Alba, the EGFP (enhanced GFP) bunny
Created in 2000 as a transgenic artwork

32. Transgenic Pigs Pass on the Transgene

33. Glofish are on sale throughout the US except in California
GloFish, originally developed in Singapore as a way to monitor water pollution
The normally black-and-silver zebrafish was turned green or red by inserting various versions of the GFP gene
Glofish are on sale throughout the US except in California
Glofish retail for about $5 per fish. Normal zebrafish cost around one tenth of the price

34. Chimeric mouse
The brown fur comes from ES cells injected into the blastocyst of an albino mouse

35. GDF8 (Myostatin) knockout mouse
normal knockout
GDF8 (Myostatin) knockout mouse
Over twice the muscle mass of a wildtype mouse

36. Clones and Cloning

37. Dolly, First Mammal Cloned From an Adult Cell
Dolly, as an adult
Dolly as a lamb with her surrogate mother

38. What Has Been Cloned So Far?
Somatic Cell Nuclear Transfer
Sheep, Goat, Mouse, Rabbit, Cattle (domestic & wild), Pig, Horse, Mule, Dog, Cat (domestic & wild), Deer
Embryo Splitting (Twinning)
Sheep, Cattle, Primate (Rhesus)

39. Cat Clone Donor Surrogate mother with clone (CC)
Out of 87 implants only CC survived to birth