Biotech II Animals Transgenics

8/2/2019 Biotech II Animals Transgenics

1/21

Biotechnology II wild animal for meat ,dairy,liether,transport

Domesticated animals fewer than 20Cattle ,sheep , goast , pigs , hoarse. 5 species .

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Dosile

Flexible in their dietary

Can grow in road maternity quickly n a herbivore diet.

Breed readily n captivity.

Have hierarchical social structurers that permit human to established.

Dominance over them.

Adapted to living in large groups.____________________________________________________________________________

Development of farm animals

Selection methods .

Potential agricultural value.

The animals adapt to thrive in a domesticated environment.


2/21

Vacccination

Freezing of semen.Testing n selection.

Use of antibiotic in feed to increase gain.

Embryo transfer.

Embryo splitting n cloning from blastomeres.

In vitro maturation in vitro fertilization of oocyte n in vitro resulting embryo.use of

hormones.

Hormone sex reversal.

Steroid administration to improve weight gain . Bovine somatotropin BST.

_____________________________________________________________________________Making transgenic animals.

Inject gene and feede fertilized egg of the desired animal using a very small glass

Needle visualized microinjection.

In approximately 1% of the injected eggs the gene is physically recombine an

eggs genome.

The new gene integrate into the genome before egg begin to ddivide.

Implanted the fertilized egg is into a mother where it fully develops transgenicanimal.


3/21

Human disease research.

Input , output , value-added traits.

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Input traits

- Increasing production efficiency.

- More efficient allergics leaks growth rates.

- Resistance to disease._______________________________________________________________________

Output traits.

Enhancing the quality of the animal product.

eaner more ee an por .

Milk leaks allergics protein

,yogurt.

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Value-added traits

Therapeutics proteinsXenotransplantation.

Models for hujman disease.


4/21

Biot in animal medication vaccine .

Medicine.

Production

Anti-bacterial compound and substitute using same antibiotion in animals.

Animal vaccine.

Rapid test kits.

_________________________________________________________________________Therapeutic in value.

Antithrombin III

Milk protein.

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Vector used for gene transferre escape n hazrd.

Environment issue.

Animal welfare issue.

The tool government department and agencies involved sufficiently well defined

n weather sufficient to cope with technologies.


5/21

Regulation agency in USA.

FDA department of agriculture,environment protection agency to regulatetransgenic animals n their prodcuts to ensure that they are safe for public

uses and environment.

Depending on the nature of the genetic modification .

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Genetically identical copies___________________________________________________________________________

Monozygotic twins

Transfer to nucleated oocyte of nuclei from blastomere,blastomere

Nucleus transfer BNT.

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Soamtic cell nuclear transfer SNT

Embryonic stem cells.

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BNT,SNT result in an exact replica of an individual animal.Mitochondria ,cytoplasmic factors. Mitochondria ,cytoplasmic factors.


6/21

Health problem.

Age prematurely.

Lack of genetic diversity ,

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Technical and application .

Germ line how transgenic animal produced?

DNA microinjection.Retrovirus mediated gene transfer.

Embryonic stem cell mediated gene transfer.

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,

their offspringAcquire the desired transgene.

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Male transgenic low

Gene transfer 0.6% transgenic pigs were injected with specific transgene.

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7/21

DNA microinjection

Transfer of the desired gene. Pronucleus.

The manipulate cell must be culture in vitro to develop a specific embryonicphase transfer to the recipient female.

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Retrovirus mediated gene transfer

Use vectors to transfer,resulting in a chimera and organism consisting

Of tissue , part of diverse genetic constitution.

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Chimera 20 generations.

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Embryonic stem cell mediated gene transferIsolation of totipotent stem cells.

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Agricultural

Breeding

Quality

Growth hormones.

Disease resistance.


8/21

Medical transplantation.

XenotransplantationTransgenic pigs then transplant organ.

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Nutritional supplement and pharmaceuticals products ,insulin , GH ,anti-clotting

factors from milk transgenic from cows sheep goats

Treat PKU phenylketonuria ,cystic fibrocis.Protein-enriched milk 2.4 gram/litre

Rosies milk alpha lactalbumin.

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uman gene erapy

Normal copy of a gene (transgene) to the genome of a person defective copies ofthe gene.

- Genetic discover A.I. produce calf and gene then promoters the growth of red

cells in human.

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Industrial application.


9/21

Industrial application.

Toxicity-sensitive produced for chemical safety testing.Microorganism engineered to rpoduced a wide variety of proteins then enzyme

speed up industry chemical reaction.

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Transgenic chicken.

Grow faster than sheep and goats .Synthesize several group of protein in the white of their eggs.

Chicken carrying an expressing foreign genes.

Infecting embryo with a viral vector copying.

uman gene or erapeu c pro e n .

Promoter sequences that will respond to the signals for making proteins(lysozyme) in egg white.

Transforming sperm and human gene and promoters checking transgenic

offspring.

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10/21

Transgenic fish

Design and construction of trasgenic DNA .Transfer gene to genome cells.

Screening for transgenic fish.

Find transgene expression and phenotype.

Study inheritance

Stable lines of transgenics.

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GH gene from large sized fish.

Human benefit protein.

Gene were introduced into medaka fish to create resistance to bacterial

pathogen.

Some commercially desirable fish and crutaceaus have been difficult to

genetically engineer because embryonic tissue is difficult to manipulate.

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11/21

Regulation of transgenic fish.

Synthetic triploid strain of fish eggs roduced from treatment of pressure

temperature shock and sex hormone the sterile triploids were tend toproduced fertile progeny established transgenic population.

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The tiny zebra fish that live in aquariums a popular laboratory animals to

produce a fluorescent red pigments.

,promoted for sale household aquarium.____________________________________________________________________________

FDA position glofish = unmodified fish.

Food safety isomers.

ene sease res s ance s o a sor ox c su s ance g.

90% food allergies ,potential for allergic reactions. Large dose it consume in

cow and uncook food

Suchi.


12/21

Eu concerns

Natural ecosystem and native species

50% of all harmful economics and environment.

66% unintentionally.

Farmed fish excape from open water contaminate native population.

Inevitable that trangenic fish will escape from aquaculture .

Inevitable that transgenic fish.Competitive advantager over native species.

Using GH gram increase growth 2-11 times.

Lead to sexual maturity, breeding,

pee roug ou na ve popu a on.

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Maturity,factor,die sooner,goes for rapid growth,shorter life expectancy.

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13/21

Transgenic cow

1. Superovulation treated

2. Fertilization

3. Gene transform by microinjection.

4. Implantation.

5. Determination of transgene expression and phenotype.

6. Study of inheritance.7. Selection of stable lines of transgenic.

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Pharmaceutical.

Disease. Resistance.

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30% from staphylococus aureus then secreting added protein. Lysostaphin

Defense bioengineering.

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Transgenic cow and blood thickener in milk.


14/21

Transgenic 3 years develop drugs.

Developed from ovaries of dead cow.

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Creat bioreactor for biopharnmaceuticals production of recombinant human growth hGH in the milk

of a cloned transgene cow at levels of up to 5 g/l

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Hormone transgenic livestock.Transgene then germline of animals.mans.

Benefit farm animals and humans.

However restricted inefficiencies methodology.

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Implantation __________________________________________________________________________________

Livestock costly.

Incorporated foreig DNA sperm mediated gene transfer SMGT.

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15/21

Phytate phosphorus pigs and chicken.

produce enzyme phytase in thesalivary glan secreted in the saliva.

100,000 untis/kg.___________________________________________________________________________________

Phosphorus in feces enzyme stable and active degrade in pancreatic. Protease.

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Transgenic dogs Alzheimers disease breakthrough. GMO animal models for alzheimers disease

Brain disorder , pharmaceutical. Industry.

Expressinng plant genes.Levels of unsaturated fatty acids > 100 fold higher

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Preadipocites derived from adipose tissue at transgenic fish cultured in vitro than intact fat tissue from

.

__________________________________________________________________________________Transgenic sheep anf goat .

Milk produced a human protein alpha antitrypain,

Treat aquatic fibrosis , 1st clone dolly the sheep.

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Human factor IX transgenic sheep produced by transfer of nuclei.

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Improve wool pronucleus microinjection the high-sulfur keratin


16/21

Golden egg a new way to make drug.

Eggs and human protein gene alter chicken.

Antibody used for passive immunization for diagnostic purposes like blood group , disease testingTherapeutic intervention.

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Monoclonal antibody mab

Fusion of B-lymphocytes immune cells of a mouse.

Myeloma cancer cell.

Hybridoma immortal cancer cell and specific antibody.

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Mab

Purification process.

es s ma recogn ze mo ecu e.

Pregnancy , fertility test.Safe

Broad range of potential targets.

High selectivty.

High affinity.

Ease of preparation._______________________________________________________________________________


17/21

Therapeutic

High therapeutic dose.

Low immunogenicity.Poor penetration in solid tumors.

Potential cross reactivity and other tumors.

High production cost.

Potential viral safety problems.

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All mab Abs were derived from rodent cells

Rodent-derived mab suffered from short half life and poor recognition of the rodent IgG-C region

Human effectors function.

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enera on o rue uman ma s ns a e an on y m e num er o arge s.

-Chimerization resulting mab 60-70%-Humanication 90-95%.

-Use phage display fully mab.

-Transgenics fully human mabs.

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18/21

The rDNA technology

Insulin from E.coliMore complex and larger gene products > insulins

Can not post translational.

Glycosylation.

Folding.

TrimmimgBiological function.

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Protein

Bioactivity erythropoietin , tissue plasminogen .activation.

Development of posttranslational processing.

Development of cell line.

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E.Coli for animal cells.

Biopharmaceutical was produced in E.coli. 86%50-60% produced in animal cells CHO Hamster.


19/21

-The development rDNA heterogenous gene expression.

-The disadvantage is the high production costs and production in cultivated bacteria , yeast cells.

- Research program have developed robust and highproducing animals host cell lines._______________________________________________________________________________

Animal cell proteins.

-Therapeutic biopharmaceuticals using animal cell technology.

-proteins used for the treatment of cardiovascular disease.

tPA ,tissue , plasminogen activation.

Cystic fibrosis Dnase , erythropoietin EPO.Viral infection , interferon and interleukin.

,multiple sclerosis , interferon-beta ,

Human growth ,GH.

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Culturing of animal cells farms also an important of gene therapy.Delivery gene to patient.

Result long lasting treatment , recombinant viruses.

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Drug discovery.

The evolution of drug discovery and development process has been revolutionized by high-throughput

Technology.

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20/21

-Chemistry.

Gemomics-based target identification.

Animals and human cells.Cell-based assays and models.

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Large scale cultivation for industrial purposes is performed in bioreactors large stainless steel tanks and

Gentle agitation and aeration to supply o2.

Cells , multiple,release protein into medium.

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Human protein.

Product sufficient amount.

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a er or e env ronmen .

__________________________________________________________________________________The vector used

Retroviral vectors.

Adenoviral vextors.

Naked , plasmid DNA.

Lipofection.

Pox viruses vector.

Vaccination viruses vectors.

Herpes simplex virus.

AAV adevo-associated viruses.


21/21

Preclinical studies to evaluate, number of gene therapy,gene therapy program for neuologic illnesses,

Autoimmune diseases, allergies, regeneration of tissues.

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Documents

Biotech II Animals Transgenics