Transgenic animals

1. By : Ravi Arora
M Pharmacy 1st year
Pharmacology
UIPS,PU

2. Transgenic Technology or
Transgenesis
Transgenesis is the technique used to
insert or transfer the foreign gene or an
altered gene into the genome of an
organsim. Thus, it is the process of
introducing an exogenous gene – called a
transgene – into a living organism so that
the organism will exhibit a new property
and transmit that property to its
offspring.

3. Transgenic Organisms
A transgenic organsim is one that carries a
foreign gene or an altered gene that has been
deliberately inserted into its genome.
Transgene
A transgene is a gene or genetic material
that has been transferred naturally or by any
of a number of genetic engineering
techniques from one organism to another.

4. Transgenesis can be facilitated by:
 Lipofection
 Plasmid vectors
 Pronuclear injection or Microinjection
 Protoplast fusion
 Transfection
 Electroporation
 Viral transformation
 Plant tissue culture

5. Lipofection
Lipofection (liposome transfection)
is a technique used to inject genetic
material into a cell by means of
liposomes, which are vesicles that can
easily merge with the cell membrane
since they are both made of a
phospholipid bilayer.

6. Microinjection
Microinjection refers to the
process of using a glass
micropipette to insert
substances at a microscopic
or borderline macroscopic
level into a single living cell.
It is a simple mechanical
process in which a needle
roughly 0.5 to 5 micrometers
in diameter penetrates the
cell membrane and/or the
nuclear envelope.

7. Embryonic Stem Cell Method
Embryo development→ blastocyst
stage
↓
8 – 16 cell cluster &
forms an early
embryo
↓
DNA introduced into ES cells
via. Transfection or microinjection
↓
ES cells carrying transgene
cultured, inserted into blastocysts,
implanted into pseudopregnant
foster mothers and allowed to
develop

8. Transfection
Transfection is the process of
deliberately introducing nucleic
acids into cells. The term is used
notably for non-viral methods in
eukaryotic cells. Transfection of
animal cells typically involves
opening transient pores or "holes"
in the cell membrane, to allow the
uptake of material. Transfection
can be carried out using calcium
phosphate, by electroporation, or
by mixing a cationic lipid with the
material to produce liposomes,
which fuse with the cell
membrane and deposit their cargo
inside.

9. Viral transformation
Viral transformation most
commonly refers to the virus-
induced malignant transformation
of an animal cell in a body or cell
culture. In molecular biology, the
term may also refer to the
transfection of DNA into a host
cell using a viral vector.

10. Choice of animal
Should breed rapidly
Produce large number of offspring
Should be easy to keep free from viral
infections
Minimum time to reach maturity
Transferred gene should be active in
the transgenic species

11. Examples
Rats: Larger, surgical
manipulation easier
Mice: Best but small in size
Baboons: Slow breeders
Pigs: Same size &
physiology as humans,
can be bred more easily,
useful for organ
transplantation

12. Applications

13. Medicine
 Insulin
 Human growth hormones
 Follistim
 Human albumin
 Monoclonal antibodies
 Antihemophilic factors
 Vaccines
 Calcitonin
 Chronic gonadotrophin
 Blood clotting factors
 Tissue plasminogen activator
 Erythropoietin
 Interferon
 Interlukins
 and many more

14. Research
 Loss of function experiments, such as in a
gene knockout experiment.
 Gain of function experiments, the logical
counterpart of knockouts.
 Tracking experiments, which seek to gain
information about the localization and
interaction of the desired protein.
 Expression studies aim to discover where and
when specific proteins are produced.

15. Oncomouse
 The OncoMouse or Harvard mouse is a type of
laboratory mouse that has been genetically
modified using modifications designed by Philip
Leder and Timothy of Harvard University to carry a
specific gene called an activated oncogene.
 The activated oncogene significantly increases the
mouse’s susceptibility to cancer, and thus makes
the mouse suitable for cancer research.
 The rights to the invention are owned by DuPont.
"OncoMouse" is a registered trademark.

16. Knockout mouse
 A knockout mouse is a genetically engineered mouse in which
one or more genes have been turned off through a targeted
mutation.
 Knockout mice are important animal models for studying the
role of genes which have been sequenced but whose functions
have not been determined. By causing a specific gene to be
inactive in the mouse, and observing any differences from
normal behavior or condition, researchers can infer its probable
function.
 Examples of research in which knockout mice have been useful
include studying and modeling different kinds of cancer,
obesity, heart disease, diabetes, arthritis, substance abuse,
anxiety, aging and Parkinson's disease.

17. Industrial
By engineering genes into bacterial plasmids it is
possible to create a biological factory that can produce
proteins and enzymes. Some genes do not work well in
bacteria, so yeast, a eukaryote, can also be used.
 supplements such as tryptophan
 chymosin in cheese making

18. Agriculture
 Tomato (Flavr Savr,Fish tomato)
 Golden rice
 Transgenic maize
 Transgenic soybean
 Bt brinjal
 Blue rose
 Knockout moss

19. Strategy
 Cloned genes injected into nucleus of a
fertilized cell
 Inoculated fertilized egg is implanted in a
receptive female
 Few offspring's carry cloned genes in all
their cells
 Animals with cloned genes in their germ
cells were bred to establish new genetic lines

20. Top Transgenic Animals
Developments

21. 1. Brainbow Mouse
 Images of brain cells in over 90
different shades.
 The colorful cells were individual
neurons and the colors helped
scientists distinguish one cell from
another.
 A team of scientists at Harvard
University were able to achieve the
specialized coloring by developing
a transgenic technology that
utilizes four fluorescent proteins
in varying combinations and
saturations.

22. 2. Spider Goat
 Transgenic goats that have the
spider silk gene inserted into
their DNA.
 These goats as a result of foreign
gene secrete a protein spider silk
in their milk.
 The substance: spider silk.
Stronger than steel and very
flexible.
 Molecular biologist and University
of Wyoming professor Randy
Lewis in collaboration with Nexia
Biotechnologies is harvesting a
protein that he believes could have
applications as far-reaching as
bulletproof vests, artificial
tendons and even clothing.

23. 3. Cancer-Resistant Mouse
 A mouse that was resistant to
spontaneous and artificially
induced tumors.
 Its inability to get cancer is due
to the insertion of a gene that
codes for a protein called Par-4.
 This protein specifically kills
cancer cells without affecting
normal cells.
 Its a tumor suppressor that
would be ideal for therapeutic
intervention strategies.

24. 4. GloFish
 World's first commercially available
transgenic animal, Yorktown
Technology's GloFish
 These zebrafish have had fluorescent
proteins extracted from jellyfish
inserted into their DNA to make them
glow green, orange, or red.
 The technology was pioneered by
Zhiyuan Gong at the National
University of Singapore.
 He was studying water pollutants by
inserting genes for proteins that would
fluoresce in the presence of certain
contaminants, but the first step was
creating a fish that fluoresced all the
time.

25. 5. Emerald Sea Slug (Elysiachlorotica)
 A slimy gastropod, emerald a sea slug
may be the first known naturally-
occurring transgenic species.
 This creature confounded researchers
of University of Maine, who
discovered that it is able to use the
chloroplasts from the algae on which
it feeds to create energy by
photosynthesis.
 Rumpho eventually found a gene in
the slug with a sequence identical to
that of an algal photosynthetic gene,
indicating that the gene may have
been naturally integrated into the sea
slug's DNA in the past.

26. 6. Aedes Mosquitoes
 Mosquitoes are the carriers for many infectious
diseases, including dengue fever, which has no
vaccine or cure.
 Company Oxitec, an offshoot of Oxford
University, is collaborating with the Malaysian
government to test a new method in which
males genetically altered to be sterile are
released into the wild.
 These infertile mosquitoes mate with females
and produce no offspring, reducing the
mosquito population.
 Though this treatment is in the early stages of
testing, but it could become a major component
of an integrated program" to fight mosquito-
mediated diseases.

27. 7. Drug-Producing Chicken
 The chicken egg could someday
produce life-saving drugs.
 Chickens are simpler and more
efficient factories than synthetic
methods for producing
pharmaceutical proteins (the
proteins are extracted from the
egg white).
 Scientists says that their next aim
is to produce an egg that contains
an antibody against a virus of the
gut. "The idea is that children
will eat the egg white and the
antibody will protect them from
developing a diarrhoeal disease."

28. 8. Mighty Mice
 The mice in the laboratory of Dr.
Richard Hanson at Case Western
Reserve University.
 He can run for 25 times as long as
normal mice at the same speed, and
they eat more and live longer.
 Bred as a tool to investigate the
metabolic enzyme PEPCK-C, which
plays a role in the generation of glucose,
the mice are genetically altered to over-
produce this enzyme.
 The mice get much of their energy
during exercise from fatty acids and
generate very little lactic acid, possibly
explaining their ability to run so long.
 Hanson believes the special abilities of
these animals make them a good model
through which to study various aspects
of aging.

29. 9. Fearless Mouse
 Gleb Shymansky at Rutgers University have
implicated a genetic basis for fear with their
transgenic mice that show deficits in their
innate anxiety and learned fear in
behavioral tests.
 They achieved these results by removing the
gene for stathmin, an enzyme that is found
in high levels in an area of the brain
implicated in emotional processing and fear.
 While these mice developed normally and
their other types of memory were normal,
they showed deficits in long-term
potentiation, a process that is necessary to
fear conditioning and may play a role in
innate fear.

30. 10. Enviropig
 As a producer of large amounts of
phosphorus, pig manure can have a
detrimental effect on the environment,
killing marine species and generating
greenhouse gases.
 To reduce this impact, Cecil Forsberg of the
University of Guelph reported in the
October 2001 issue of Nature Biotechnology
that his lab had bred a transgenic pig that
produces phytase, an enzyme that aids in
the breakdown of phosphorus, in its saliva.
 These pigs produce up to 75 percent less
phosphorus in their waste.
 FDA approved, the Enviropig for
consumption in the United States as early
as this year.

31. 11.Designer baby
 The colloquial term designer baby refers to a baby
whose genetic makeup has been artificially selected
by genetic engineering combined with in vitro
fertilisation to ensure the presence or absence of
particular genes or characteristics.
 The term is derived by comparison with "designer
clothing".
 It implies the ultimate commodification of children
and is therefore usually used pejoratively to signal
opposition to such use of reprogenetics.

32. Thank You