The document discusses transgenic animals, which are genetically modified to carry foreign genes for various purposes, including medical applications, agriculture, and research models for human diseases. It outlines methods for producing transgenic animals, such as nuclear microinjection and retrovirus-mediated gene transfer, and provides examples like the superfish and Dolly the sheep. The document emphasizes the potential benefits and applications of transgenic technology across multiple fields, while noting that regulatory approval for consumption is still lacking.

1. TRANSGENIC
ANIMALS
Submitted To :- Submitted By :-
Govind Singh Sumita
Hooda
Department of Pharmaceutical Sciences,
Maharshi Dayanand University, Rohtak-124001

2. Content:-
 Transgenic Animals
 Definition,
 Purpose,
 Some Species In Which Transgenic Animals Have Been
Produced,
 Production Of Transgenic Animals
 Some Example Of Transgenic Animal,
 Transgenic Animals Model Of Human Disease,
 Application Of Transgenic Animal,
 Reference.

3. TRANSGENIC ANIMALS
Definition:-
A transgenic animal is one that carries a foreign
gene that has been deliberately inserted into it
genome.
The first transgenic animal was a ‘Mouse’, but
now or some year later it is followed by
Rabbit, Pigs, Sheep, Cattle.
Currently, no transgenic animal or animal
product is approved by the FDA or USDA for
human consumption.

4. PURPOSE
 Medical applications
 In the basic biological study,
 Xenotransplantation,
 In nutritional, supplements and
pharmaceutical,
 As model of human disease,
Fig:-Vacanti Mouse

5. PURPOSE
 In industry
 in toxicology,
 in biotechnology.
Fig:-See-Through Frog

6. PURPOSE
 Agriculture and aquaculture application
 Breeding,
 Quality,
 Disease resistance.
Fig:-Enviropig

7. Some Species In Which
Transgenic Animals Have Been
Produced
Mice (Mus musculus) Gordon et al. (1980), Joyner and Sedivy
(2000)
Rats (Rattus rattus) Hamra et al. (2002), Kato et al. (2004),
Hirabayashi et al. (2005), Agca et al. (2008)
Rabbits (Oryctolagus
cuniculus)
Fan and Watanabe (2003)
Sheep (Ovis aries) McCreath et al. (2000), Denning and Priddle
(2003), Wheeler (2003)
Pigs (Sus domestica) Lai et al. (2002), Houdebine (2009), Kragh et
al. (2009)
Cattle (Bos taurus) Donovan et al. (2005), Richt et al. (2007),
Houdebine (2009)
Goats (Capra hircus) Wheeler (2003), Houdebine (2009)
Dogs (Canis familiaris) Hong et al. (2009)
Marmosets (Callithrix jacchus) Sasaki et al. (2009)

8. Production Of Transgenic
Animals
Three basic methods for producing
transgenic animal.
 Nuclear Microinjection
 Embryonic Stem Cells
 Retrovirus Mediated Gene Transfer

9. Nuclear Microinjection
MICRO
NEEDLE
OOCYTE
PIPP
ET

10. Nuclear Microinjection
Once the suitable gene is available, the standard
scenario for the creation of a transgenic animal by
nuclear microinjection is as follows:-
 The transgene is injected into fertilized cell (above
fig). Just after fertilization, the egg contains its
original female nucleus from the successful sperm.
These two pronuclei will soon fused together.
Before this happen, the DNA is injected into the
male pronucleus, which is larger and better target
for microinjection.
 Continued….

11. Nuclear Microinjection
 Nucleus microinjection requires specialized
equipment and great skills.{the success rate varies
from 5% to 40% among various laboratories}
 The egg is kept in culture during the first few
divisions of embryonic development.
 The engineered embryos are then implanted into
the womb of a female animal, the forest mother.
They develop into embryos and if it goes well, into
new born animals.
 Continued…

12. Nuclear Microinjection
 Some of the baby animal will have the transgene
stably integrated into their chromosomes known as
founder animal.
 In other, the process fail and transgene is lost.
 A male and female mated to form a new line of
animals carrying two copies of the transgene.
 The founder animals contain only a single copy of
the transgene on one chromosome and are
heterozygous for the transgene.
 Continued….

13. Nuclear Microinjection
 When to such animals are bred together,25% of the
progeny will get two copies of the transgene and
will be homozygous,25% will get zero copies and
the remaining 50% will get one copy.
 Homozygous are most useful because if they are
further interbred, all of their descendants will get
two copies of the transgene.

14. Embryonic Stem Cell

15. Embryonic Stem Cell
 Stem cells are the precursor cells to particular tissue
of body.
 Embryonic stem cells are derived from bastocyt, a
very early stage of the embryo and retain the ability
to develop into any body tissue including the
germline.
 Embryonic stem cells can be cultured and DNA can
be introduced as for any cultured cell line.
 Continued…………

16. Embryonic Stem Cell
 For successful creation of transgenic animal,
embryonic stem cell mus be maintained under
conditions that avoid differentiation.
 Engineered embryonic stem cells are then inserted
into the center cavity of the blastocyt stage. This
process creates a mixed embryo and the result in an
animal that is genetic chimera consisting of some
transgenic tissue and other that are normal.

17. Retrovirus Mediated Gene
Transfer
 Eggs are infected prior to fertilization
 Virus integrates into one of the chromosomes

18. Retrovirus Mediated Gene
Transfer
 The most useful vector for the purpose of the gene isolation
are those that lend themselves to the production of the
libraries consisting of overlapping fragments of genomic DNA
ideally encompassing the entire genome several times.
 Ex:- Bacteriophage λ genomic library of 106 viruses each
containing on overage 20kb of DNA represent 6-7 copies of
the entire mouse genome and the probability that each gene is
represented is very high.
 Retroviruses can be used for the transfer of the foreign genes
into animal genomes.
 Continued…………

19. Retrovirus Mediated Gene
Transfer
 This can best be done at 4-16 cell stage embryos. However it can be
done up to midgestion, but with incomplete infections. i.e. low
infectivity rate.
 Immediately following injection, the retrovirus produces a DNA
copy of its RNA genome using its reverse transcriptase.
 Completion of this process requires that host cell undergoes the s-
phase of the cell cycle. Therefore retroviruses effectively transduce
only mitotically active cells.
 Modification to the retrovirus frequently consist of removal of
structural gene, such as gag, pol and eve, which support viral
particle formation.
 Continued…………

20. Retrovirus Mediated Gene
Transfer
 Additionally, most retrovirus and complementary lines are
ecotropic in that they infect only rodent, such as rats & mice,
and rodent cell lines rather than humans.
 The DNA copy of the viral genome or provirus, integrates
randomly into the host cell genome, usually without deletions
or rearrangements.
 Because integration is not by way of homologous
recombination, this method is not used effectively for site
directed mutagenesis.
 Very high rates of gene transfer are achieved with the use of
retrovirus.
 Continued…………

21. Retrovirus Mediated Gene
Transfer
 Vector used for such purposes:-
Origin Insert size
range(kb)
Multicopy plasmids Multicopy plasmids Up to 20kb
Lambda vectors Bacteriophage λ Up to 30kb
Cosmid Bacteriophage λ Up to 40kb
P1 artificial chrom BacteriophageP1 80-90kb
Bacterial artificial
chrom
Large bacteria plasmid 100-300kb(f-factor)
Yeast chrom [YAC] Yeast chromosome 100-1000+ kb
+ means indefinite.

22. Retrovirus Mediated Gene
Transfer
 DISADVANTAGE:-
 Low copy number integration.
 Additional steps required for produce retrovirus.
 Limitation on size of the foreign DNA insert( usually
9 to 15 kb transferred).
 Potential for undesired genetic recombination that may
be alter the retrovirus.
 Possible interference by integrated retroviral sequences
in transgene expression.

23. Some Example Of Transgenic
Animal
 Growth hormone:- Superfish
 Increased growth and size.
 Growth hormone gene inserted
into fertilized egg.
 Transgenic salmon grows about 10-11
times faster than normal fish.

24. Some Example Of Transgenic
Animal
 Glofish:- genetically modification

25. GLOFISH
 The GloFish is a patented brand of genetically modified (GM) fluorescent
zebrafish with bright red, green, and orange fluorescent color. The original
zebrafish from which the GloFish was developed measures three
centimeters long and has gold and dark blue stripes. In 1999, Dr. Zhiyuan
Gong and his colleagues at the National University of Singapore were
working with a gene called green fluorescent protein (GFP), originally
extracted from a jellyfish, that naturally produced bright green
bioluminescence. They inserted the gene into a zebrafish embryo, allowing
it to integrate into the zebrafish’s genome, which caused the fish to be
brightly fluorescent under both natural white light and ultraviolet light.
Their goal was to develop a fish that could detect pollution by selectively
fluorescing in the presence of environmental toxins.
 It is the first genetically modified animal to become
publicly available as a pet.

26. Some Example Of Transgenic
Animal
 Dolly the sheep

27. DOLLY THE SHEEP
 Not so cool or disturbing enough but dolly would hit this list for
sure since she was the first ever cloned animal which means that she
was produced from a single microscopic cell from a single parent
(who hadn’t mated of-course). Cloning techniques might be used
widely now in some part of worlds for food but dolly remains
remarkable in being the first mammal to be cloned from an adult
somatic cell, using the process of nuclear transfer. Normally off-
springs are a result of interaction of sex cells but in case of dolly’s
birth, sex cells weren’t involved. She was cloned by Ian Wilmut,
Keith Campbell and colleagues at the Roslin Institute near
Edinburgh in Scotland. She was born on 5 July 1996 and she lived
until the age of six. She has been called “the world’s most famous
sheep” by sources including BBC News and Scientific American. To
good, dolly was fertile and produced 6 lambs in total. She died in
2003, living about half as long as a typical sheep. She developed a
lung disease common in older sheep.

28. Some Example Of Transgenic
Animal
 Ruppy:- Ruppy (short for Ruby Puppy) is a cloned
beagle from South Korea who glows red under
ultraviolet light. Ruppy was created in 2009 by a group
of scientists in South Korea, led by Byeong-Chun Lee.
The dog was cloned using viral transfection of
fibroblasts cells with a
protein that expresses the
red fluorescent gene.

29. Some Example Of Transgenic
Animal
 See-Through Frog

30. SEE-THROUGH FROG
• Dissecting animals for science has sparked
controversies worldwide, even prompting some
companies to create computer simulations as
cruelty-free alternatives. For high school students
everywhere, this revealing amphibian may be a cut
above regular frogs. That’s because the see-through
frog does not require dissection to see its organs,
blood vessels, and eggs. You can see through the
skin how organs grow, how cancer starts and
develops. It’s a miracle of genetic engineering and
surely a cool mutant gift to students.

31. Some Example Of Transgenic
Animal
 Enviropig
 A genetically engineered pig approved for limited
production which produces 65% less phosphorous in
animal waste thus very environmental friendly.

32. Transgenic Animals Model Of Human
Disease
1. Human Immunodeficiency Virus/ Acquired
Immunodeficiency Syndrome (HIV/AIDS):
 Tg26 HIVAN Mouse Model was the first
transgenic model developed in 1991 for HIV.
 These transgenic animals can express HIV-1
proteins develop symptoms and immune
deficiencies similar to the manifestations of AIDS
in humans.
 Other models are AIDS Mouse and Smart Mouse.

33. Transgenic Animals Model Of Human
Disease
2. Alzheimer’s disease:
 Mouse models like PDAPP mice, Tg2576 mice,
TAU transgenic mouse models like ALZ7 mice,
7TauTg mice.
 Immunization of Amyloid precursor protein A42
in transgenic mice showed that vaccination
against Alzheimer’s disease could have potential
as a therapeutic approach.

34. Transgenic Animals Model Of Human
Disease
3. Cardiovascular disease:
 Transgenic models of heart failure and hypertrophy
like Gene overexpression of Calmodulin, Gene
mutation of alpha cardiac myosin heavy chain and
Knockout gene model of transforming growth
factor are developed.
 Mutation of the ApoE gene that is critical for
uptake of chylomicrons and very low density
lipoprotein particles, results in a model that
develops atherosclerotic lesions histologically
similar to those found in humans.

35. Transgenic Animals Model Of Human
Disease
4. Diabetes Mellitus:
 A transgenic mouse model that expressed
Insulin Dependent Diabetes Mellitus by
inserting a viral gene in the animal egg stage is
also developed.
 There are other models like beta receptor
knockout mouse, uncoupling protein (UCP1)
knockout mouse , acute and chronic models for
the evaluation of anti-diabetic agents.

36. Transgenic Animals Model Of Human
Disease
5. Angiogenesis:-
 Mouse models of angiogenesis, arterial stenosis,
atherosclerosis, thrombosis, thrombolysis and
bleeding addresses techniques to evaluate
vascular development.
 With the help of angiogenesis transgenic animal
models inhibitors are identified which act on
specific mechanisms of angiogenesis.

37. Transgenic Animals Model Of Human
Disease
6. Cancer diseases:
 Oncomouse was first transgenic animal to be
patented.
 Mechanisms for tumor progression and
metastasis via E-cadherin, and other adhesion
molecules is possible by various transgenic
knockout models.

38. Transgenic Animals Model Of Human
Disease
7. Some other examples of transgenic
animal model
Speies Protein product Potential indication(s)
Cow Collagen Burns bone fracture
Goat Monoclonal antibodies Colon cancer
Pig Factor ix and factor viii Hemophilia
Rabbits Insulin like growth
factor
Wound healing
Sheep Protein C Deficiency, adjunct of tPA
Goat LAtPA2 Myocardial infarct, pulmonary
embolism
Pig Human hemoglobin Blood replacement for
transfusion

39. Application of transgenic
animals
 Normal Physiology and Development:-Transgenic animals
can be specifically designed to allow the study of how genes
are regulated and how they affect the normal functioning of
the body and its development. For example, the study of
complex factors involved in growth such as insulin likes
growth factors.
 Study of Disease:- Many transgenic animals are designed to
increase the understanding of that how genes contribute to the
development of diseases such as cancer, cystic fibrosis,
rheumatoid arthritis and Alzheimer.

40. Application of transgenic
animals
 Biological Products:-Human disease can be treated by
medicines that contain biological products.
(a) Transgenic animals that produce useful biological products
can be created by the introduction of the portion of the
DNA or genes that codes for a particular product such as
human protein (alpha-1-antitrypsin) which is used to treat
emphysema.
b) Similar attempts are being made for the treatment of
phenylketonuria (PKU) and cystic fibrosis. For example,
the first transgenic cow Rosie produced human protein
enriched milk (2.4 g/L) in 1997. The milk contained the
human alpha lactalbumin and was nutritionally a more
balanced product for human babies than natural cow milk.

41. Application of transgenic
animals
 Vaccine Safety:-Transgenic mice are being used in testing the
safety of vaccines before they are used in humans, e.g., polio
vaccine.
These animals are also used for the toxicity or safety testing
procedures. If found reliable and successful they could replace
the use of monkeys in order to test the safety of batches of the
vaccine.
 Chemical Safety Testing:- Transgenic animals are made to
carry the genes, which make them more sensitive to the toxic
substances than the non-transgenic ones. They are then
exposed to toxic substances and effects are studied. This is
known as toxicity/safety testing.

42. Conclusion
 Transgenic technology is a field that is under constant
evolution.
 Many transgenic animal have been successfully created
for a Varity of purposes and prospects are enormous.
 It hold great potential in many fields including
agriculture, medicine and industry.
 With proper research and careful use the transgenic
animals can go a long way in solving several problems
for which science doesn't have a solution till now.

43. REFERENCES
 Eppeudrof A G, Sonja V, “Application Of Transgenic Animals”,
Hamburg, Germany June 2006, 1-9.
 Bogle T. R. , Kunkulol R. R. , Baig M. S. , More S. Y., “Transgenic
Animals And Their Application In Medicine”, “International
Journal Of Medical Research And Health Sciences”, Vol 2 issue 1
jan-mar 2013, 107-116.
 Clark D. P. , Pazdernik N. J. ,“Transgenic Animals”,
“Biotechnology Applying The Genetic Revolution”, Academic
Press Cell, 2nd Edition 2016, 425-456
 Crommelin D. J. A. , Sindelar R. D. , Pharmaceutical Biotechnology
Fundammental and Application, bernd meibolim, editor springer, 4th
edition, 2012, pg. no. 220-233.
 Periera F. R., “Animal Biotechnology” , “ Wisdom Press, New
Delhi, Editior 2013, pg. no. 58-83.

44. REFERENCES
 http://www.smashinglists.com/10-mutant-and-genetically-modified-
organisms/2/
 Khan F. A. , “biotechnology fundamentals”, “ CRC Press, 1st Edition
2012,pg. no. 220-223.
 Sosa M. A., Gasperi R.D., Elder G. A. , “Animal transgenesis: an
overview”, Springer- verlag 2009.
 Sasidhara R. , “ Animal Biotechnology” MJP Publisher, Chennai 2006, 1st
Edition, pg. no. 133-164.
 Louis-Marie Houdebine, “Methods to Generate Transgenic Animals”
 Louis-Marie Houdebine, “The methods to generate transgenic animals and
to control transgene expression”, “Journal of Biotechnology” 98 (2002)
145–160