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DEVELOPEMENT AND
USES OF TRANSGENIC
ANIMALS
BY
M.NAGAPRADHEESH(2K21/MSBT/18)
II M.Sc., BIO TECHNOLOGY
ANIMAL BIOTECHNOLOGY
DEFINITIONS :
TRANSGENIC ANIMALS (OR) GENETICALLY
MODIFIED ORGANISMS (GMO)
• Genetically Modified Animals (GMO) and the animals which contains, gene of interest
or foreign DNA is called as Transgenic Animals.
• Genetically modified animals are animals that have been genetically modified for a
variety of purposes including producing drugs, enhancing yields, increasing resistance
to disease, etc. The vast majority of genetically modified animals are at the research
stage while the number close to entering the market remains small.
• The process of genetically engineering animals is a slow, tedious, and expensive
process. The development of the CRISPR-Cas9 gene editing system has effectively
halved the amount of time needed to develop genetically modified animals.
HISTORY AND DEVELOPEMENTS OF TRANSGENIC ANIMALS:
• 1907-Ross Granville and Harrison working at John Hopkins Medical School,for the
first time established the methodology of animal tissue culture by culturing the frog
cells.
• 1966-Dr.Ian Wilmut and colleagues carried out nuclear transfer to make xerox animals
from parents.This is now called as Animal Cloning.
• 1974-Rudolf Jenisch created a Transgenic Mouse by introducing foreign DNA into its
embryo making world’s first Transgenic Animal.
• 1982-Thilly and colleagues established the scaling up process for the large scale
culture of animal cells.
• 1985-The first transgenic livestock were produced.(By Gene Knockout Method)
• 1989-Knockout Mices are produced from one gene (or) one cell.
• 1996-”DOLLY” THE SHEEP. The firstever trangenic animal was cloned and Identified by
Dr.Ian Wilmut at Roselin Institute,Scotland.
• 1998- Thomas and Gearhart isolate and culture human embryonic stem cells for research
purpose.
• 2003-The first genetically modified animal to be commercialised was the GloFish, a Zebra
fish with a fluorescent gene added that allows it to glow in the dark under ultraviolet
light.It was released to the US market.
• 2015-The first genetically modified fish to be approved for food use was AquAdvantage
Salmon.
• 2017-Chinese scientists announced they used CRISPR gene editing technology to create of
a line of pigs with better body temperature regulation, resulting in about 24% less body fat
than type.
DIFFERENT METHODS USED FOR DEVELOPEMENT
OF TRANSGENIC ANIMALS:
• The transgenic animals can be produced by three
different methods:
1. MICROINJECTION METHOD
2.RETRO VIRAL METHOD
3.EMBROYONIC STEM CELL METHOD.
1.MICROINJECTION METHOD:
■Large foreign genes can be transferred to the receipient cells by
microinjection.
• A female mice superovulated by injecting pregnant mar’s serum and Human chronic
gonototropin in two days interval.
• The Mice produce about 35 eggs
• The female mice is mated with a male
• Fertilised eggs are taken from the oviduct and kept in BSS(Balanced Salt Solution).
• The eggs are visualized under the microscope to pick up eggs having male and female pro
nucleus
• The foreign gene microinjected into the male pro nucleus of the egg.
• About 25-30 microinjected eggs are implanted in the uterus of the female mice.
• Then the mice is mated with a vasectomised male.
• These mating bring up a suitable condition for the further developement of eggs in uterus.
• The mother will give birth to transgenic pups.
• And the transgenic pups will contain the desired characterstics of the desired gene.
2.RETRO VIRAL METHOD:
■Small foreign gene less than 8kg bp can be transferred to
receipient cells,through retro-virus vectors.
■Murine Leukemia Virus(MULV) is the most common retro-
virus being used to introduce genes into animal cells.
• The Mice cell line is infected with MULV.
• RNA of MULV produces ds-DNA by reverse transcription.
• The Pro viral DNA is isolated from the cell line.
• The foreign gene is linked with a neomycin resistance gene using DNA Ligase
• The foreign DNA is inserted into the Pro viral DNA using a proper restriction enzyme and DNA
Ligase
• The resulting hybrid DNA is called as r-DNA(recombinant DNA).
• The r-DNA is introduced into a mice cell-line by calcium phosphate mediated gene transfer.
• The cell line is then infected with helper Murine Leukemia Virus that can produce viral
capsid.
• The r-DNA produce recombinant viral RNA,later gets packed into the viral capsid to form
recombinant murine leukemia virus
• Sperm and eggs were collected and fertilised in invitro condition to form Zygotes.
• After 8-celled stage the mice embryos are infected with recombinant murine leukemia
virus.
• The Embryos are cultured in the medium containing neomycin in order to screen the
recombinants.
• The living embryos are implanted into the uterus of the serrogate mother.
• Then the female is mated with a vasectomised male mice for the further developement of
the egg in uterus.
• After the pregnancy the serrogate mother will produce the transgenic animals with the
desired characters.
3.EMBRYONIC STEM CELL METHOD:
■Mice embryos at blastocyst stage are collected from
pregnant female mice.
■Embryonic Stem Cells present in the blastocyst are
isolated and kept in BSS.
• The desired foreign gene is linked a neomycin resistance gene.
• The embryonic stem cells which contains the r-DNA are transferred to a medium
containing neomycin
• Recombinant embryonic stem cells can alone grow in the medium and the others die.
• The recombinant embryonic stem cells are microinjected into fresh Blastocyst taken
from mice.
• The Microinjected Blastocyst are implanted into the female mice and mated
Vasectomised male
• After the pregnancy period,the mother will produce transgenic pups with the desired
characters.
APPLICATIONS OF TRANSGENIC ANIMALS:
1. Transgenic mice have been used as human disease models for
AIDS,cancer,diabetes,Alzheimer’s diseases etc...
2. Super ovulated Mouse-The transgenic mouse with growth harmone gene
will have a increased wait,which can be treated for meat production.
3. Animal farming,production of pharamaceutically valuable human proteins
farm the milk of transgenic animals.
4. Transgenic Cow with K-Caesin protein gene will produce milk rich in K-
Caesin with milk is suitable for chese making.
5. Transgenic Cattle with lactice gene of interest produce milk which is pour
in lactose.This milk is likely for lactose intolerant people.
6. Mammary Glands of the Transgenic Sheep serves as bioreactions to
produce semi-proteins of high therapeutic value.
7. Transgenic Sheep is used for wool production.
8. Transgenic creates with TPA (Tissue Plasminogen Actinator) gene will produce TPA in
milk which is used to treat dissolve blood clots.
9.Xenotransplantation-Transplantation of animal organs in human transgenic pigs with
certain genes from human-immune system will have human proteins.
10.Trangenic Virus will produce Bacterial resistance chicken.
11.Transgenic Chicken that produces eggs which are lower in fat in cholesterol.
12.Transgenic Chicken with high protein in that Meat.
13.Transgenic Fish was made by introducing human growth harmone which grow faster
than the normal fish.
14.Transgenic Mosquitoes with gene for endotoxin of Plasmodium Vivax will produce
toxin of malarial parasite in their saliva when the mosquito inject the brain human blood
immune system will produce antibodies against the toxin.
ADVANTAGES OF TRANSGENIC ANIMALS:
(a) Gene requires certain cellular mechanism to help for the production of protein.
The
animals used for transgenic purpose naturally carry the mechanism needed to
produce
complex protein. Theses mechanism is absent in cell culture.
(b) Expression through cell culture or bacterial culture requires constant monitoring
and
sampling.
(c) The isolation and purification of expressed protein in conventional method is
more
difficult than purifying proteins from an animal’s milk or body fluid.
(d) It is more cost effective as the product is efficiently passed through milk with an
average yield of 53% and with 99% purity.
(e) It has been estimated that transgenic animal can produce in its lifetime $100 to
$200 million worth of pharmaceuticals.
DISADVANTAGES OF TRANSGENIC ANIMALS:
(a) Transgenic animal project is extremely expensive.
(b) Generation of transgenic animals are also expensive, because of long gestation
period, litter size and higher maintenance cost of the recipient animals.
(c) There may be high mortality rate and other deleterious effects on animals used by
researchers to create transgenic breeds. It has been observed that transgenic pigs
having
enhanced growth rate and efficient feed conversion exhibit reduced reproductive
performance and may suffer from arthritis and dermatitis etc.
(d) Large number of recipients is required for embryo transfer because of low
transgenesis rate.
(e) Transgenic foods have been produced and offer better productivity in terms of both
yield and quantity. However, there are some apprehension about the safety of
transgenic
food
REFERENCES:
• WEBSITES:
☆https//:www.thebiotech notes.com/Transgenic
animals
☆https//:www.microbenotes.in/Animal cell culture.
• BOOKS:
TITLE:TEXTBOOK OF BIOTECHNOLOGY
AUTHOR:H.K.DAS
EDITION:4th EDITION
PUBLICATION:WILEY INDIA.
Transgenic Animals developement and uses(M.NAGAPRADHEESH).pptx

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Transgenic Animals developement and uses(M.NAGAPRADHEESH).pptx

  • 1. DEVELOPEMENT AND USES OF TRANSGENIC ANIMALS BY M.NAGAPRADHEESH(2K21/MSBT/18) II M.Sc., BIO TECHNOLOGY ANIMAL BIOTECHNOLOGY
  • 2. DEFINITIONS : TRANSGENIC ANIMALS (OR) GENETICALLY MODIFIED ORGANISMS (GMO) • Genetically Modified Animals (GMO) and the animals which contains, gene of interest or foreign DNA is called as Transgenic Animals. • Genetically modified animals are animals that have been genetically modified for a variety of purposes including producing drugs, enhancing yields, increasing resistance to disease, etc. The vast majority of genetically modified animals are at the research stage while the number close to entering the market remains small. • The process of genetically engineering animals is a slow, tedious, and expensive process. The development of the CRISPR-Cas9 gene editing system has effectively halved the amount of time needed to develop genetically modified animals.
  • 3. HISTORY AND DEVELOPEMENTS OF TRANSGENIC ANIMALS: • 1907-Ross Granville and Harrison working at John Hopkins Medical School,for the first time established the methodology of animal tissue culture by culturing the frog cells. • 1966-Dr.Ian Wilmut and colleagues carried out nuclear transfer to make xerox animals from parents.This is now called as Animal Cloning. • 1974-Rudolf Jenisch created a Transgenic Mouse by introducing foreign DNA into its embryo making world’s first Transgenic Animal. • 1982-Thilly and colleagues established the scaling up process for the large scale culture of animal cells. • 1985-The first transgenic livestock were produced.(By Gene Knockout Method) • 1989-Knockout Mices are produced from one gene (or) one cell.
  • 4. • 1996-”DOLLY” THE SHEEP. The firstever trangenic animal was cloned and Identified by Dr.Ian Wilmut at Roselin Institute,Scotland. • 1998- Thomas and Gearhart isolate and culture human embryonic stem cells for research purpose. • 2003-The first genetically modified animal to be commercialised was the GloFish, a Zebra fish with a fluorescent gene added that allows it to glow in the dark under ultraviolet light.It was released to the US market. • 2015-The first genetically modified fish to be approved for food use was AquAdvantage Salmon. • 2017-Chinese scientists announced they used CRISPR gene editing technology to create of a line of pigs with better body temperature regulation, resulting in about 24% less body fat than type.
  • 5. DIFFERENT METHODS USED FOR DEVELOPEMENT OF TRANSGENIC ANIMALS: • The transgenic animals can be produced by three different methods: 1. MICROINJECTION METHOD 2.RETRO VIRAL METHOD 3.EMBROYONIC STEM CELL METHOD.
  • 6. 1.MICROINJECTION METHOD: ■Large foreign genes can be transferred to the receipient cells by microinjection. • A female mice superovulated by injecting pregnant mar’s serum and Human chronic gonototropin in two days interval. • The Mice produce about 35 eggs • The female mice is mated with a male • Fertilised eggs are taken from the oviduct and kept in BSS(Balanced Salt Solution). • The eggs are visualized under the microscope to pick up eggs having male and female pro nucleus • The foreign gene microinjected into the male pro nucleus of the egg.
  • 7. • About 25-30 microinjected eggs are implanted in the uterus of the female mice. • Then the mice is mated with a vasectomised male. • These mating bring up a suitable condition for the further developement of eggs in uterus. • The mother will give birth to transgenic pups. • And the transgenic pups will contain the desired characterstics of the desired gene.
  • 8. 2.RETRO VIRAL METHOD: ■Small foreign gene less than 8kg bp can be transferred to receipient cells,through retro-virus vectors. ■Murine Leukemia Virus(MULV) is the most common retro- virus being used to introduce genes into animal cells. • The Mice cell line is infected with MULV. • RNA of MULV produces ds-DNA by reverse transcription. • The Pro viral DNA is isolated from the cell line. • The foreign gene is linked with a neomycin resistance gene using DNA Ligase • The foreign DNA is inserted into the Pro viral DNA using a proper restriction enzyme and DNA Ligase • The resulting hybrid DNA is called as r-DNA(recombinant DNA).
  • 9. • The r-DNA is introduced into a mice cell-line by calcium phosphate mediated gene transfer. • The cell line is then infected with helper Murine Leukemia Virus that can produce viral capsid. • The r-DNA produce recombinant viral RNA,later gets packed into the viral capsid to form recombinant murine leukemia virus • Sperm and eggs were collected and fertilised in invitro condition to form Zygotes. • After 8-celled stage the mice embryos are infected with recombinant murine leukemia virus. • The Embryos are cultured in the medium containing neomycin in order to screen the recombinants. • The living embryos are implanted into the uterus of the serrogate mother. • Then the female is mated with a vasectomised male mice for the further developement of the egg in uterus.
  • 10. • After the pregnancy the serrogate mother will produce the transgenic animals with the desired characters.
  • 11. 3.EMBRYONIC STEM CELL METHOD: ■Mice embryos at blastocyst stage are collected from pregnant female mice. ■Embryonic Stem Cells present in the blastocyst are isolated and kept in BSS. • The desired foreign gene is linked a neomycin resistance gene. • The embryonic stem cells which contains the r-DNA are transferred to a medium containing neomycin • Recombinant embryonic stem cells can alone grow in the medium and the others die. • The recombinant embryonic stem cells are microinjected into fresh Blastocyst taken from mice.
  • 12. • The Microinjected Blastocyst are implanted into the female mice and mated Vasectomised male • After the pregnancy period,the mother will produce transgenic pups with the desired characters.
  • 13. APPLICATIONS OF TRANSGENIC ANIMALS: 1. Transgenic mice have been used as human disease models for AIDS,cancer,diabetes,Alzheimer’s diseases etc... 2. Super ovulated Mouse-The transgenic mouse with growth harmone gene will have a increased wait,which can be treated for meat production. 3. Animal farming,production of pharamaceutically valuable human proteins farm the milk of transgenic animals. 4. Transgenic Cow with K-Caesin protein gene will produce milk rich in K- Caesin with milk is suitable for chese making. 5. Transgenic Cattle with lactice gene of interest produce milk which is pour in lactose.This milk is likely for lactose intolerant people. 6. Mammary Glands of the Transgenic Sheep serves as bioreactions to produce semi-proteins of high therapeutic value. 7. Transgenic Sheep is used for wool production.
  • 14. 8. Transgenic creates with TPA (Tissue Plasminogen Actinator) gene will produce TPA in milk which is used to treat dissolve blood clots. 9.Xenotransplantation-Transplantation of animal organs in human transgenic pigs with certain genes from human-immune system will have human proteins. 10.Trangenic Virus will produce Bacterial resistance chicken. 11.Transgenic Chicken that produces eggs which are lower in fat in cholesterol. 12.Transgenic Chicken with high protein in that Meat. 13.Transgenic Fish was made by introducing human growth harmone which grow faster than the normal fish. 14.Transgenic Mosquitoes with gene for endotoxin of Plasmodium Vivax will produce toxin of malarial parasite in their saliva when the mosquito inject the brain human blood immune system will produce antibodies against the toxin.
  • 15. ADVANTAGES OF TRANSGENIC ANIMALS: (a) Gene requires certain cellular mechanism to help for the production of protein. The animals used for transgenic purpose naturally carry the mechanism needed to produce complex protein. Theses mechanism is absent in cell culture. (b) Expression through cell culture or bacterial culture requires constant monitoring and sampling. (c) The isolation and purification of expressed protein in conventional method is more difficult than purifying proteins from an animal’s milk or body fluid. (d) It is more cost effective as the product is efficiently passed through milk with an average yield of 53% and with 99% purity. (e) It has been estimated that transgenic animal can produce in its lifetime $100 to $200 million worth of pharmaceuticals.
  • 16. DISADVANTAGES OF TRANSGENIC ANIMALS: (a) Transgenic animal project is extremely expensive. (b) Generation of transgenic animals are also expensive, because of long gestation period, litter size and higher maintenance cost of the recipient animals. (c) There may be high mortality rate and other deleterious effects on animals used by researchers to create transgenic breeds. It has been observed that transgenic pigs having enhanced growth rate and efficient feed conversion exhibit reduced reproductive performance and may suffer from arthritis and dermatitis etc. (d) Large number of recipients is required for embryo transfer because of low transgenesis rate. (e) Transgenic foods have been produced and offer better productivity in terms of both yield and quantity. However, there are some apprehension about the safety of transgenic food
  • 17. REFERENCES: • WEBSITES: ☆https//:www.thebiotech notes.com/Transgenic animals ☆https//:www.microbenotes.in/Animal cell culture. • BOOKS: TITLE:TEXTBOOK OF BIOTECHNOLOGY AUTHOR:H.K.DAS EDITION:4th EDITION PUBLICATION:WILEY INDIA.