1. Transgenic Organisms

2. Definitions: Genetically modified organisms (GMO’s):
-Organisms whose genes have been altered using genetic engineering techniques.
Transgenic organisms
- Most GMO’s are transgenic organisms… they have received genes from a different organism.
Cry genes (produce Bt toxin) from Bacillus thurengensis (Bt) put into peanut plant - European corn borer larvae
Genetically engineered sheep with the human gene responsible for the production of alpha-1-antitrypsin (AAT) transferred into their DNA. When the AAT is eventually extracted from their milk, it will be used as therapy for humans deficient in AAT. The deficiency causes emphysema (a breathing disorder) in approximately 100,000 people in the western world.
Ex. A mouse is given a gene from a human. The mouse is a transgenic GMO.
Trans- ; across (across species in this case)

3. How Do They Do It?

5. GMO’s at home: GloFish Zebra danio
1. Zebra danio was genetically engineered with a gene from sea coral that causes the fish to glow in the presence of environmental toxins.
2. Gene was inserted into the embryo of the fish.
3. First GMO available as a pet.

6. (jellyfish, phylum cnidaria)
GMO’s in research:
GFP (green fluorescent protein)
GFP Mice
1. Gene from a jellyfish (Aequorea victoria) that codes for GFP was inserted into the embryos of mice.
Aequorea victoria
(jellyfish, phylum cnidaria)

7. GMO’s in research: 1. GFP is used in cellular and molecular biology.
GFP (green fluorescent protein) – a reporter protein
1. GFP is used in cellular and molecular biology.
2. You can attach this protein to any other protein you want making it a reporter protein.
- It “reports” to you where the protein is going since it emits green light (similar to radioactivity in that sense)

8. GMO’s in research:
- GFP has been attached to a protein called MFD, which is found in peroxisomes.
- Those little green dots are peroxisomes…
- You can track any protein you want…in a single cell or an entire organism

9. 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
                                                        

10. 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

11. As water temperature drops the GH gene is turned on
The transgene used to increase growth utilizes an antifreeze protein promoter connected to the GH cDNA
Antifreeze promoter from pout
As water temperature drops the GH gene is turned on
The fish continue to grow when normally they would not

12. Concerns if these ‘supersized’ transgenic fish got loose:
Transgenic fish are farm-raised, isolated from wild stocks
But even during farming of wild type fish, escapes happen frequently (~14 million/yr)
What would happen if a large number of transgenic escaped & started breeding with wild fish?

13. In experiments, transgenic males mated 3x more frequently than the smaller wild males
Offspring of transgenic males lived <70% as long as wild males
Could lead to a decline of the wild fish population & endanger a species as whole

14. Antifreeze Proteins (AFP)
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

15. This is a genetically modified strain of malaria-resistant mosquito which has been created successfully by a scientist.
The creation of mosquitoes with green fluorescent testicles will help curb the spread of malaria-carrying mosquitoes.

16. Animal Bioreactors
“Pharming”

17. GMO medicine: E. Coli with the human insulin gene
Cry genes (produce Bt toxin) from Bacillus thurengensis (Bt) put into peanut plant - European corn borer larvae
Genetically engineered sheep with the human gene responsible for the production of alpha-1-antitrypsin (AAT) transferred into their DNA. When the AAT is eventually extracted from their milk, it will be used as therapy for humans deficient in AAT. The deficiency causes emphysema (a breathing disorder) in approximately 100,000 people in the western world.
Alpha 1-Antitrypsin or α1-antitrypsin (A1AT) is a glycoprotein and generally known as serum trypsin inhibitor. Alpha 1- antitrypsin is also referred to as alpha-1 proteinase inhibitor (A1PI) because it is a serine protease inhibitor (serpin), inhibiting a wide variety of proteases.[1] It protects tissues from enzymes of inflammatory cells, especially elastase, and has a reference range in blood of gram/liter (in US the reference range is generally expressed as mg/dL or micromoles), but the concentration can rise manyfold upon acute inflammation.[2] In its absence, elastase is free to break down elastin, which contributes to the elasticity of the lungs, resulting in respiratory complications such as emphysema, or COPD (chronic obstructive pulmonary disease) in adults and cirrhosis in adults or children.
- Insulin is made using the bacterium E. coli.
- The human gene coding for insulin is inserted into E. coli, which will then make insulin for us (we will see how this is done shortly)…

18. GMO medicine: AAT Sheep
Cry genes (produce Bt toxin) from Bacillus thurengensis (Bt) put into peanut plant - European corn borer larvae
Genetically engineered sheep with the human gene responsible for the production of alpha-1-antitrypsin (AAT) transferred into their DNA. When the AAT is eventually extracted from their milk, it will be used as therapy for humans deficient in AAT. The deficiency causes emphysema (a breathing disorder) in approximately 100,000 people in the western world.
Alpha 1-Antitrypsin or α1-antitrypsin (A1AT) is a glycoprotein and generally known as serum trypsin inhibitor. Alpha 1- antitrypsin is also referred to as alpha-1 proteinase inhibitor (A1PI) because it is a serine protease inhibitor (serpin), inhibiting a wide variety of proteases.[1] It protects tissues from enzymes of inflammatory cells, especially elastase, and has a reference range in blood of gram/liter (in US the reference range is generally expressed as mg/dL or micromoles), but the concentration can rise manyfold upon acute inflammation.[2] In its absence, elastase is free to break down elastin, which contributes to the elasticity of the lungs, resulting in respiratory complications such as emphysema, or COPD (chronic obstructive pulmonary disease) in adults and cirrhosis in adults or children.
Genetically engineered sheep with the human gene for alpha-1-antitrypsin (AAT).
AAT is extracted from their milk and used to treat humans deficient in AAT, which is one cause of emphysema (a breathing disorder) in approximately 100,000 people in the western world.

19. GMO’s and medicine:
Transgenic cows can make serum albumin for treatment of burns and severe trauma to tissue

20. Other Types of Transgenic Animals

21. Spider silk
Spider’s silk is 5 times stronger than a thread of steel of the same thickness 3 times stronger than Kevlar (carbon fibre)
Spiders are carnivores and cannot be raised
Transgenic animals can produce the spider protein
Aculepeira ceropegia
a European orbweb spider
© P. Billiet
© 2010 Paul Billiet ODWS

22. Spider silk
The gene for spider silk protein was isolated from the golden orb weaver (Nephila clavipes)
It is a fibrous protein so it has a very repetitive gene sequence
Nephila clavipes © C. Frank Starmer
© 2010 Paul Billiet ODWS

23. Web-based kid
Spider goat © Sean O'Neill
Gene (plus regulator genes) inserted into goat mammary gland cells
Genetically transformed cells could be made to secrete spider silk protein
Transformed goat cells fused to enucleated oocyte
Genetically transformed goat embryos produced
© 2010 Paul Billiet ODWS

24. Applications Super strong surgery thread Artificial ligaments
Light bullet proof body armor
Biodegradable fishing line
© 2010 Paul Billiet ODWS

25. ANDi, the first transgenic primate born in Jan 2000
                                                
ANDi, the first transgenic primate born in Jan 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

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

27. Researchers transfer DNA from the long-vanished Tasmanian tiger into a mouse. The finding shows how lost information about species from the past can be retrieved and also provides a glimpse into how long-gone creatures may someday get a second chance at life.

28. Mouse “Knock-out” Technology
Gene Targeting

29. Knock-out technology allows for the specific loss of a gene in mice
Allows for the function of the KO’d gene to be deduced from the defects seen in the mice
Can be used to mimic some disease
Unlike traditional transgenics, the trangene is targeted to a specific site in the DNA of the mouse

30. Mouse Knock-outs require embryonic stem (ES) cells
These are derived from the inner cell mass (ICM) of a blastocyst (the ICM is what will become the fetus)
ES cells are pluripotent meaning they can become all the different cell types found in an adult

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

32. Some Examples of Knockout Organisms

33. p27 knockout mouse is bigger than the control
This is not due to obesity, but the skeletal structure is increased in size (everything about the mouse is larger)

34. GDF8 (Myostatin) knockout mouse
normal knockout
GDF8 (Myostatin) knockout mouse
Over twice the muscle mass of a wild type mouse

35. Naturally Occurring GDF8 Mutants

36. Conclusion
- We can basically move any gene(s) between members of a species or between any species.
- We can also alter the genes to our liking (GFP tagged proteins) before inserting them into embryos.
Cry genes (produce Bt toxin) from Bacillus thurengensis (Bt) put into peanut plant - European corn borer larvae
Genetically engineered sheep with the human gene responsible for the production of alpha-1-antitrypsin (AAT) transferred into their DNA. When the AAT is eventually extracted from their milk, it will be used as therapy for humans deficient in AAT. The deficiency causes emphysema (a breathing disorder) in approximately 100,000 people in the western world.
Is all of this genetic engineering positive or negative?