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ANIMAL CLONING
PROCEDURES AND APPLICATION
Animal Cloning Procedures
and Application
Elfren F. Celestino Jr.
INTRODUCTION
• Purpose of genetic engineering is direct
• Genetic engineering is a technique used for direct genetic manipulation of DNA sequences.
modification of organisms using recombination of DNA.
• These techniques involve the capacity to
• Identify, replicate, modify and transfer the genetic material of isolate, cut and transfer specific DNA pieces,
cells, tissues or complete organisms. corresponding to specific genes.
• Cloning is the creation of an organism that is
an exact genetic copy of another. The
following 3 types of cloning technologies are:
(1) recombinant DNA technology
or DNA cloning,
(2) reproductive cloning, and
• Dolly (1997-2003), a Finn Dorsett ewe, was (3) therapeutic cloning.
the first mammal to have been successfully
cloned from an adult cell.
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DNA CLONING Steps in Synthesizing a Recombinant Protein:
• Procedure of isolating a defined DNA sequence
and obtaining multiple copies of it in vivo. • isolation of a gene of interest.
• The objectives of Recombinant DNA :
• The gene is then inserted into a vector
– Identifying genes
• A vector is a piece of DNA that is capable of
– Isolating genes
independent growth (bacterial plasmids and
– Modifying genes viral phages).
– Re-expressing genes in other hosts or
organisms
Restriction Enzymes are Enzymes That Cut DNA Only at
Particular Sequences
The enzyme EcoRI cutting DNA at its recognition sequence
Different restriction enzymes have different
recognition sequences.
• a ligation procedure is employed whereby
the gene of interest is inserted into a vector.
• The vector is linearised by means of
restriction enzymes, and incubated with the
gene of interest under appropriate conditions
with an enzyme called DNA ligase
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• The gene of interest (foreign DNA) is • Once the vector is isolated in large quantities,
integrated into the plasmid or phage it can be introduced into the desired host
recombinant DNA.
cells
• Before introducing the vector containing the
foreign DNA into host cells to express the • The host cells will then synthesize the foreign
protein, it must be cloned. protein from the recombinant DNA.
• Cloning is necessary to produce • When the cells are grown in vast quantities,
numerous copies of the DNA since the the foreign or recombinant protein can be
initial supply is inadequate to insert isolated and purified in large amounts.
into host cells.
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Cloning Vectors Plasmid Cloning Vectors
• Plasmids are circular,
• A vector is used to amplify a single molecule double-stranded DNA
of DNA into many copes. A DNA fragment molecules
must be inserted into a cloning vector. • They can replicate
• A cloning vector is a DNA molecule that has independently of the
host cell.
an origin of replication and is capable of
• Plasmids have an origin
replicating in a bacterial cell. of replication, antibiotic
• Most vectors are genetically engineered resistance genes as
plasmids or phages. markers, and several
unique restriction sites.
Plasmids are Used to Replicate a Recombinant DNA REPRODUCTIVE CLONING
• Plasmids are small circles of DNA found in • "somatic cell nuclear transfer" (SCNT) to
bacteria. create animals that are genetically identical.
• Plasmids replicate independently • This process entails the transfer of a nucleus
from a donor adult cell to an egg which has
• Pieces of foreign DNA can be added within a no nucleus.
plasmid to create a recombinant plasmid. • If the egg begins to divide normally it is
transferred into the uterus of the surrogate
mother
Applications of Reproductive Cloning THERAPEUTIC CLONING
• Somatic cell nuclear transfer can also be used
• Would allow infertile parents to have a child to create a clonal embryo.
that is genetically related to one of them
• Therapeutic cloning (embryo cloning) is the
• Would allow a couple in which one partner production of human embryos for use in
has a dominant genetically transmitted research.
disease to have a child genetically related to
• The goal of this process is not to create
one of them without risk of the disease’s
being transmitted cloned human beings, but rather to harvest
stem cells that can be used to study human
development and to treat disease.
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Applications of Therapeutic Cloning SOMATIC CELL NUCLEAR
TRANSFER
• Would allow the production of embryos with
specific genetic traits for us in research on • somatic cells are taken from the animal that
that trait. is going to be cloned.
• The nucleus of somatic cell (the donor cell) is
• Would allow the production of embryos from removed and the rest of the cell discarded.
which stem cells could be derived. • At the same time, the nucleus of an egg cell is
removed.
• The nucleus of the somatic cell is then
inserted into the enucleated egg cell. • The division of the egg cell follows the same
process that would occur if the egg was
• The egg is then stimulated to grow by fertilised by sperm during natural
numerous stimulants which activates the reproduction.
reconstructed embryo to divide and grow. • The cell division continues for 5 days until a
The two cells were fused by blastomere forms
electrical shock (called • Blastomer can be implanted into a uterus of a
“electrofusion”). female to create a whole organism
• Blastomeres can also be used as a source of
stem cells.
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Health aspect
• The stresses placed on both the egg cell and – Many clones are born with defects
the introduced nucleus are enormous,
leading to a high loss in resulting cells. • kidney problems
• For example, Dolly the sheep was born after • diabetes
277 eggs were used for SCNT, which created • crippling disabilities
29 viable embryos. Only three of these
embryos survived until birth, and only one
survived to adulthood.
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• In SCNT, not all of the donor cell's genetic Benefits of Genetically Engineered
information is transferred, as the donor cell's Animals
mitochondria that contain their own
mitochondrial DNA are left behind.
• The resulting hybrid cells retain those
• Used to develop new medical treatments
mitochondrial structures which originally
belonged to the egg. • Improve our food supply
• As a consequence, clones such as Dolly that • Enhance our understanding of all animals,
are born from SCNT are not perfect copies of including humans
the donor of the nucleus.
Benefits of Cloning Increased disease prevention and
detection
• Reduce variability of responses of a
population being used to test new drugs New diagnostic tests for diseases in
companion animals, livestock, and
• Preservation of endangered species
poultry are being developed to
• Reduce time to produce new breeds of farm diagnose animal diseases, and
animals assess the health state of animals.
Treatments may involve the production
of more specific animal vaccines
APPLICATION
b. Pet Cloning
a. Conservation Biology
Below are Baba Ganoush and Tabouli,
Noah, the first cloned endangered species. created by Genetic Savings & Clone, Inc.
Noah is a gaur, a species endangered by
hunting and habitat loss.
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d. Disease Models
c. Developmental and Basic Research
HIV normally will
In January 2001, at only cause disease
the Oregon Regional in humans and
chimpanzees. By
Primate Research knocking out a
Center created ANDi mouse’s immune
system and using
by inserting into his human stem cells to
germline a jellyfish create a human
gene for immune system, HIV
mice are susceptible
bioluminescence. The to AIDS.
gene is present, but
inactive.
e. Xenotransplantation f. Growth Hormone Genes
Xenotransplantation is the transplantation of
living cells, tissues or organs from one species
The Beltsville
to another such as from pigs to humans hogs had a human
gene for growth
In 1984, Baby Fae hormone inserted
received the heart of a
baboon and survived into their genome.
for three weeks.
g. Pharming - is the production of
pharmaceuticals in animals engineered
to contain a foreign, drug-producing gene.
Tracy has been
genetically
engineered
by PPL Therapeutics
to produce alpha1-
antitrypsin (AAT),
useful for treating
emphysema.
Dolly and her surrogate mother
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The first cloned horse and her surrogate mother
Five genetically identical cloned pigs.
CopyCat Tetra
In January 2000,
researchers at the
Oregon Health
Sciences University
reported that they
had produced
Researchers are interested in using cloned cats Tetra.
in AIDS research, since feline AIDS is a good
model for human AIDS.
