 Transgenesis is the process of introducing
an exogenous gene—called a transgene—
into a living organism so that it will exhibit a
new property and transmit that property to its
offspring.
 A transgenic animal is one whose genome
has been changed to carry genes from
other species.
1. DNA Microinjection
o Transfer of a desired gene into the pronucleus of a reproductive
cell.
o The manipulated cell, which first must be cultured in vitro.
o Then transferred to the recipient female.
2. Retrovirus-Mediated Gene Transfer
 Retroviruses used as vectors to transfer genetic material into the
host cell, resulting in a chimera, an organism consisting of tissues
or parts of diverse genetic constitution.
3. Embryonic Stem Cell-Mediated Gene Transfer
This method involves:
 Isolation of totipotent stem cells from embryos.
 The desired gene is inserted into these cells.
 cells containing the desired DNA are incorporated into the host’s
embryo, resulting in a chimeric animal.
 Unlike the other two methods, which require live transgenic
offspring to test for the presence of the desired transgene, this
method allows testing for transgenes at the cell stage.
 Bacillus thuringiensis produce this
parasporal crystal.
 Active form- binding in gut epithelial cells
of insects.
 Leakage of cellular ATP, thereby
metabolism ceases.
 Insect become dehydrated & finally dies.
INSECT RESISTANCE
 Movement protein ex- transgenic tobacco
plants.
 Transmission protein.
 Antisense RNA’s.
 Inserting a transgene into
chromosomes, a new function is
introduced while producing
transgenic animals.
 The chromosomes carrying the
target gene (4 exons) with
flanking sequences is subjected
to homologous recombination
with a vector carrying a
selectable marker gene.
 The homologous recombination
results in gene knockout
(ie.,)disruption of the target gene.
 Resistance to viral, bacterial and coccidial diseases.
 Better feed efficiency.
 Lower fat and cholesterol levels in eggs.
 Better meat quality.
 Eggs with pharmaceutical proteins in them.
Transgenic tilapia
 which is modified with pig growth-hormone, has
three times larger than their non-transgenic
siblings.
 Tilapia genetically modified could also serve as a
source of islet cells for transplantation to human
subjects.
Transgenic Zebra Fish:
 The tiny zebra fish that lives in aquariums, was
genetically modified to produce a fluorescent red
pigment, and is being promoted for sale as a
household aquarium pet, the “goldfish”.
Transgenic Fish may be better used for the following
purposes:
(1) For increasing fish production to meet the growing due to
demand of food due to increase in world population.
(2) For production of pharmaceutical and other industrial products.
(3) For development of transgenic native glow fish varieties for
aquarium.
(4) As fish biosensors for monitoring aquatic pollution.
(5) For isolation of genes, promoters and synthesis of effective
gene constructs.
(6) For researches in embryonic stem cells and in-vitro embryo
production.
(7) For production of anti-freeze protein.
1. Microinjection
2. Transposons
3. Lentiviral vector
4. DNA transfer via
sperm
5. Pluripotent cells
6. Somatic cell nuclear
transfer.
 Isolation of gene of interest
 Construction of transgene
 Transfering gene into bovine cells
 Selection for positive cells
 Confirming the cow is transgenic
 Production of therapeutic proteins
The main advantages of making therapeutic proteins in
transgenic cows are:
capacity – large amounts of protein are produced
cost – transgenesis is cheaper than mammalian cell culture
scale – it is easy to scale production up or down to meet
demand.
function – proteins are folded and fully functional
harvesting – proteins can readily be purified from milk.
 Increased milk production
 Increased wool production
 Increased growth rate
 Improved disease resistance
 Increased muscle mass
 Improved wool quality
 Improved nutritional quality.
 Production of human haemoglobin in the
blood of transgenic pigs- trauma patients.
 Transgenic pigs can be used as better
models for retinitis pigmentosa, CVD,
diabetes etc.
 Cell tracking
Thank you

Genetic engineering

  • 2.
     Transgenesis isthe process of introducing an exogenous gene—called a transgene— into a living organism so that it will exhibit a new property and transmit that property to its offspring.  A transgenic animal is one whose genome has been changed to carry genes from other species.
  • 3.
    1. DNA Microinjection oTransfer of a desired gene into the pronucleus of a reproductive cell. o The manipulated cell, which first must be cultured in vitro. o Then transferred to the recipient female. 2. Retrovirus-Mediated Gene Transfer  Retroviruses used as vectors to transfer genetic material into the host cell, resulting in a chimera, an organism consisting of tissues or parts of diverse genetic constitution. 3. Embryonic Stem Cell-Mediated Gene Transfer This method involves:  Isolation of totipotent stem cells from embryos.  The desired gene is inserted into these cells.  cells containing the desired DNA are incorporated into the host’s embryo, resulting in a chimeric animal.  Unlike the other two methods, which require live transgenic offspring to test for the presence of the desired transgene, this method allows testing for transgenes at the cell stage.
  • 4.
     Bacillus thuringiensisproduce this parasporal crystal.  Active form- binding in gut epithelial cells of insects.  Leakage of cellular ATP, thereby metabolism ceases.  Insect become dehydrated & finally dies. INSECT RESISTANCE
  • 5.
     Movement proteinex- transgenic tobacco plants.  Transmission protein.  Antisense RNA’s.
  • 8.
     Inserting atransgene into chromosomes, a new function is introduced while producing transgenic animals.  The chromosomes carrying the target gene (4 exons) with flanking sequences is subjected to homologous recombination with a vector carrying a selectable marker gene.  The homologous recombination results in gene knockout (ie.,)disruption of the target gene.
  • 11.
     Resistance toviral, bacterial and coccidial diseases.  Better feed efficiency.  Lower fat and cholesterol levels in eggs.  Better meat quality.  Eggs with pharmaceutical proteins in them.
  • 12.
    Transgenic tilapia  whichis modified with pig growth-hormone, has three times larger than their non-transgenic siblings.  Tilapia genetically modified could also serve as a source of islet cells for transplantation to human subjects. Transgenic Zebra Fish:  The tiny zebra fish that lives in aquariums, was genetically modified to produce a fluorescent red pigment, and is being promoted for sale as a household aquarium pet, the “goldfish”.
  • 13.
    Transgenic Fish maybe better used for the following purposes: (1) For increasing fish production to meet the growing due to demand of food due to increase in world population. (2) For production of pharmaceutical and other industrial products. (3) For development of transgenic native glow fish varieties for aquarium. (4) As fish biosensors for monitoring aquatic pollution. (5) For isolation of genes, promoters and synthesis of effective gene constructs. (6) For researches in embryonic stem cells and in-vitro embryo production. (7) For production of anti-freeze protein.
  • 14.
    1. Microinjection 2. Transposons 3.Lentiviral vector 4. DNA transfer via sperm 5. Pluripotent cells 6. Somatic cell nuclear transfer.
  • 15.
     Isolation ofgene of interest  Construction of transgene  Transfering gene into bovine cells  Selection for positive cells  Confirming the cow is transgenic
  • 16.
     Production oftherapeutic proteins The main advantages of making therapeutic proteins in transgenic cows are: capacity – large amounts of protein are produced cost – transgenesis is cheaper than mammalian cell culture scale – it is easy to scale production up or down to meet demand. function – proteins are folded and fully functional harvesting – proteins can readily be purified from milk.  Increased milk production  Increased wool production
  • 18.
     Increased growthrate  Improved disease resistance  Increased muscle mass  Improved wool quality  Improved nutritional quality.
  • 20.
     Production ofhuman haemoglobin in the blood of transgenic pigs- trauma patients.  Transgenic pigs can be used as better models for retinitis pigmentosa, CVD, diabetes etc.  Cell tracking
  • 21.