A transgenic animal is one that has had foreign DNA inserted into its genome. The first transgenic animal was a mouse created in 1982 by inserting a human growth hormone gene. Transgenic animals are created through pronuclear microinjection or stem cell methods. They have applications in medicine, agriculture, and industry. However, some argue that transgenic technology raises ethical issues.

1. By,
DAMARIS BENNY DANIEL
II Msc. Zoology

2.  A transgenic animal is one that carries a foreign gene that
has been deliberately inserted into its genome.
 Transgenesis is the process by which mixing up of genes
takes place.
 Foreign genes are inserted into the germ line of the
animal, so it can be transmitted to the progeny.
 Transgenic technology has led to the development of
fishes, live stock and other animals with altered genetic
profiles which are useful to mankind.

3.  First transgenic animal was a ‘Supermouse’ created by
Ralph Brinster (U Pennsylvania) and Richard Palmiter
(University of Washington) in 1982.
 It was created by inserting a human growth hormone
gene in mouse genome.
 The offspring was much larger than the parents.
 Mouse – common transgenic expt.
 Other animals include pig, goat, cow, sheep, fish etc.

4.  Step 1 – Construction of a transgene
◦ Transgene made of 3 parts:
 Pomoter
 Gene to be expressed
 Termination sequence
 Step 2 – Introduction of foreign gene into the animal
◦ Pronuclear microinjection method
◦ Embryonic stem cell method.

5.  A female animal is superovulated and eggs are collected.
 The eggs are fertilized in vitro.
 The transgene containing solution
is injected into the male pronucleus
using a micropipette.
 Eggs with the transgenes are kept overnight in an incubator
to develop to a 2 cell stage.
 The eggs are then implanted into the uterus of a pseudo -
pregnant female (female which has been mated with a
vasectimized male the previous night)

6.  Transgenic animals can be created by manipulating embryonic
stem cells.
 ES cells are obtained from the inner cell mass of a blastocyst.
 Transgene is incorporated into the ES cell by
◦ Microinjection
◦ By a retro virus
◦ By electroporation
 Transgenic stem cells are grown in vitro.
 Then they are inserted into a blastocyst and implanted into a
host’s uterus to grow normally.

8.  Step 3: Screening for transgenic positives
◦ Transgenic progenies are screened by PCR to examine the site
of incorporation of the gene
◦ Some transgenes may not be expressed if integrated into a
transcriptionally inactive site.
 Step 4: Further animal breeding is done to obtain
maximal expression.
◦ Heterozygous offsprings are mated to form homozygous
strains.

10.  Multiple insertion – too much proteins
 Insertion into an essential gene – lethality
 Insertion into a gene leading to gene silencing
 Insertion into a different area can affect the gene
regulation

12.  Superfish
◦ Increased growth and size
◦ Growth hormone gene inserted
into fertilized egg.
◦ Transgenic salmon grows about
10 – 11 times faster than normal
fish.
 Glo fish
◦ GM freshwater zebra fish (Danio
rerio)
◦ Produce by integrating a
fluorescent protein gene from
jelly fish into embryo of fish.

13.  Alzheimer’s mouse
◦ In the brain of Alzheimer’s patients, dead
nerve cells are entangled in a protein
called amyloid.
◦ Mouse made by introducing amyloid
precursor gene into fertilized egg of mice.
 Oncomouse
◦ Mouse model to study cancer
◦ Made by inserting activated oncogenes.
 Smart mouse
◦ Biological model engineered to overexpress NR2B receptor in the
synaptic pathway.
◦ This makes the mice learn faster like juveniles throughout their
lives.

14.  Enviro pig
◦ Pigs have trouble fully digesting a
compound known as phytate found in many cereal grains used
to feed them.
◦ Transgenic pigs are created by introducing phytase gene
of E.coli.
◦ Phytase enzyme is thus produced in the salvary gland of pig
◦ It degrades indigestible phytate with the release of phosphate
that is readily digested by pigs.
 Pig for organ transplant
◦ Pigs with human genes, in order to decrease the chance of
organ rejection by human body.

15.  Bioreactors whose cells have been engineered to synthesis
marketable proteins.
 More economical than producing desired protein in cell culture.
 Transgenic cattle
◦ Transgenic cows are made to produce proteins lactoferrin and
interferons in their milk.
◦ Prion free cows resistant to mad cow disease.
 Transgenic sheep
◦ For good quality wool production.
 Transgenic goat
◦ Goats that could express tissue plasminogen
activator, anti thrombin III, spider silk etc in milk.

16.  Alba, the EGFP (Enhanced Green Flurescent
protein) bunny
 Created in 2000 as a transgenic artwork.
 ANDi was the first transgenic monkey, born in 2000.
 “ANDi” stands for “inserted DNA” spelled backwards.
 An engineered virus was used to insert the harmless
gene for green fluorescence protein (GFP) into
ANDi’s rhesus genome.
 ANDi proves that transgenic primates can be
created, and can express a foreign gene delivered
into their genome.

17.  Medical importance
◦ Disease model
◦ Bioreactors for pharmaceuticals
◦ Xenotransplantation
 Agricultural importance
◦ Disease resistant animals
◦ For improving quality and quantity of milk, meat, eggs and wool
production
 Industrial importance
◦ Toxicity sensitive transgenic animals to test chemicals.
◦ Spider silk in milk of goat

18.  Blurring the lines between species by creating transgenic
combinations.
 There may be health risks associated with transgenics.
 There may be long term effects on the environment when
transgenic animals are released into the field.
 Various bioethicist argue that it is wrong to create animals that
would suffer as a result of genetic alteration.

19.  Transgenic goats engineered to produce human breast
milk ( Journal Transgenic Research, August 2012)
◦ University of California scientists created the transgenic goats by
transferring human genes for breast milk enzymes and proteins
into goat embryos.
◦ Produce 60 percent of the lysozyme and lactoferrin found in
human mother’s milk.
◦ For babies of mothers who aren’t present, or can’t nurse them,
milk from these transgenic goats could provide the next-best
alternative.
 Bioluminescent transgenic mouse model for study
mammary gland tumour development. (PLoS ONE, aug 2012)
◦ Research done by Cancer Biology and Therapeutics Group, Ireland

20.  Transgenic technology is a field that is under constant evolution.
 Many transgenic animals have been successfully created for a
variety of purposes, and the prospects are enormous.
 It holds 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.

21.  Satyanarayana U, Biotechnology (2010), 1st edition, Books and
allied (P) Ltd, Kolkata.
 Channarayappa, Molecular Biotechnology – Principles and
Practices (2006), 1st edition, University Press Pvt Ltd, Hyderabad.
 Smith E John, Biotechnology (1996), 3rd edition, Cambridge
University Press
 http://dsc.discovery.com/technology/tech-10/genetic-
engineering/10-transgenic-animals-03.html
 www.biomedcentral.com/1471-2407/12/209
 http://news.e-healthsource.com/index.php?p=news1&id=534197