Animal cloning

What is a Clone?

• Clone–a group of two or more

individuals with identical genetic
(nuclear) makeup, derived by asexual
reproduction, from a single common
parent or ancestor.

• Clones are NOT the exact copies

WHOLE ANIMAL CLONING
 Methods of animal cloning
• Nuclear transfer

• Embryo splitting

• Blastomere separation
 Nuclear transfer

Nucleus of an unfertilized egg is

replaced with the nucleus of an
donor cell.
 Embryo splitting

•To split an embryo into

pieces, each forming a
new embryo (usually, it
is to split a 2 to 8
celled embryo before
the cells differentiate).

•It is actually a simple

form of cloning
 Blastomere separation

outer coating is
removed from 4-
celled embryo, causing
separation of
individual embryo cells
(bastomeres), which
are then cultured in
vitro to develop
embryo clones
Cloning by embryo splitting
Nuclear transfer using blastomers or cultured cells as nuclear donors
Somatic Cell Nuclear Transfer
(SCNT or NTSC)
 Chronology of advances in animal
cloning
– Hans Spemann(1938) proposed use of nuclear transfer to clone an
organism.This idea originates from theoretical considerations of an
experimental method which would allow to determine whether a loss of
genetic material occurs during cell differentiation. If occurs –cloning
would not be possible.

– frog Rena pipiens(1952) Robert Briggsand Thomas Kingcloned northern

leopard frogsfirst animal cloning

– frog Xenopus leavis(1958, 1962) John B. Gurdonannounced that he had

cloned South African frogs using the nucleus of fully differentiated
adult intestinal cells. This demonstrated that cells' genetic potential do
not diminish as the cell became specialized.

• carp(1963) controversial information

• sheep (1996) the first mammal cloned: Dolly
– mouse (1998) Wakayama, Perry and Yanagimachi50 mice from adult cells
(1997-1998)
 Dolly

• Dolly(July 5, 1996 –February 14, 2003) -the first mammal to be cloned from an adult
somatic cell, using the process of nuclear transfer.
• She was cloned by Ian Wilmut, Keith Campbelland colleagues at the Roslin Institute in
Edinburgh, Scotland
• 277 oocytes were fused with cultured mammary gland cells: of these 29 reached
morula/blastula stage, and were transferred into surrogate mothers leading to 13
pregnancies, but only one live birth.

• During the Dolly`s lifetime, a lot of information on cloned organism development was
collected. It was noticed that the age of the genetic material of cloned organisms is
identical with the donor`s age. It means that biologically Dolly was 6 years old at her birth
(age of the donor sheep). This causes that clone suffers from genetic disorders of
mature/old-age donor and shows reduced disease resistance.
• At the age of five, Dolly developed arthritis.

• On February 14 , 2003 (at the age of 6 years and 7 months; the life-span for sheep: 11-12
years) Dolly was euthanized because of a progressive lung disease.
• She was bred with a Welsh Mountain ram and produced six lambs in total.
• Dolly was named after the country western singer Dolly Parton.
 Chronology of advances in
animal cloning

Monkey Rhesus:from Tetra(female,

2000)embryo splitting
pig(5 piglets Millie, Christa, Alexis,
Carrel, and Dotcom from one sow;
Scotland-2000)
cow: Alpha andBeta
(female,2001)
 cat: CopyCat ”CC”
a shorthaired calico (female, 2001)
 Chronology of advances in
animal cloning

• mouse: in2002HochedlingerandJaenischcloned
mice from T lymphocytes: They showed that
the genome of cloned individuals contains the
same rearrangement of T lymphocyte receptor
as the original population of lymphocytes. This
finding is commonly considered to be the
strongest prove for organism cloning from
differentiated cells.

• rabbit: (2003) in France and South Korea

 Chronology of advances in
animal cloning
• mule: Idaho Gem(male, May 2003)

• and Utah Pioneer(male, July 2003)

• deer: Dewey(2003)

• horse: Prometeaan Arabian

• thoroughbred (female, 2003)

• rat: Ralph(male, 2003)

• fruit flies,Drosophila(2004)

• HUMAN (2004) Shin Yong Moon`s groupThey obtained pluripotent stem cells
derived from cloned human blastocysts.

• dog: SnuppyAfghan hound (April 2005)

• wolf:grey wolves Snuwolfand Snuwolffy(2007), South Korea
Why develop cloning technology?
Potential applications of animal cloning

• Rapid multiplication of desired livestock

• Phenotypic evaluation and selection
• Animal conservation
• Transgenic applications
• Human cell‐based therapy
 Potential applications of animal
cloning

• Rapid multiplication of desired livestock

rapid spread of superior genotypes
• production of large number of
preferred sires for natural breeding
(alternative to artificial insemination)
• rapid response to market changes
A 3-year-old Friesian dairy cow, and her
genetically identical cloned calves.
Potential applications of animal
cloning

– Cloning for phenotypic evaluation and

selection to monitor effect of different
environmental conditions on phenotypes of
different lines of cloned animals enhance
genetic progress by increasing the accuracy
of selection
 Potential applications of animal
cloning
• Animal conservation cryobanking of somatic cells
from rare and endangered birds and animals
• “insurance policy” against further losses of
diversity or possible extinction of wildlife
• to preserve endangered indigenous breeds of
livestock adapted to particular environments

• Lady, the first surviving cow of the Enderby Island

breed
• (a rare breed of New Zealand origin, adapted to the
harsh sub antarctic environment), and Elsie her
genetic duplicate.
Potential applications of animal
cloning
 Potential applications of animal
cloning
– Human cell-based therapy to treat disorders in tissues
that neither repair nor replace themselves: diabetes,
muscular dystrophy, spinal cord injury, certain cancers,
various neurological disorders

– healthy cells from patient could be used to produce

therapeutic tissue for transplantation: insulin-producing
cells pancreatic cells for diabetes
– dopamine-producing nerve cells for Parkinson disease

• this may also be associated with gene therapy to correct a

genetic defect in the somatic cells (e.g. muscular
dystrophy)
Potential applications of animal
cloning
– Transgenic applications: cultured cells
can be genetically modified in the lab
– the modifications can be checked prior to
any animal work is conducted
– desired genetically modified cell lines may
then be used in nuclear transfer
– nuclear transfer approach is more efficient
than pronuclear injection method
 Uses of animal cloning for
transgenic applications
• Pharmaceuticals production of medically important molecules in milk, e.g.:Human α-
1 antytrypsin for emphysema and cystic fibrosis treatment

• Human antithrombin III to regulate blood clotting during surgery

• Nutraceuticals improved milk products, e.g.bovine milk proteins replaced with

human equivalents
• different fatty acid profiles (CLA -conjugated linoleic acid isomers with
anticancerogenic activity)

• Agricultural production traits genomics projects will identify target genes

(influencing livestock production traits):to improve feed conversion efficiency,
growth rate, muscle composition
• confer disease and pest resitance

• superior milk for cheese production increased casein and reduced β-lactoglobulin
 Uses of animal cloning for
transgenic applications
• Xenotransplantation Shortages of human organs and tissues
for transplantation
– Pigs are currently used for organ production
– Availability
– Similar anatomy, organ size and physiology
– Problematic: disease (need to generate pathogen-free herds)

– Modification of antigenes to reduce transplant rejection

– Inactivation of α-1,3-galactosyl transferase gene

– •Livestock as human genetic disease models Sheep as

alternative to mice models could be generated as superior research
models:Cystic fibrosis
– Huntington disease
 Views on Cloning

• In the debate over cloning there are those that

feel that advances gained from cloning outweigh
any social dilemmas.

• There are those that feel that cloning is wrong

on a fundamental moral level and would produce
scientific and social problems.
 Pros on Cloning

• Cloning will improve the overall quality of science

and life.
• Cloning might produce greater understanding of the
causes of miscarriages.
• Cloning experience may add to the increased
understanding of genetics.
• Damage to nervous system could be treated
through therapeutic cloning.
• Human cloning could be used for parents who risk
passing a genetic defect to a child.
• Human cloning will allow a woman to have one set of
identical twins.
 Cons on Cloning

• Those that do not agree with cloning feel that is an

affront to religious societies.
• Cloning may reduce genetic variability.
• Cloning may cause people to settle for the best existing
animals.
• Cloning is currently an expensive process.
• There is a risk of disease transfer.
• Any research into human cloning would eventually need to
be tested on humans.
• Human cloning might be used to create a “perfect human”.
• Human cloning might have a detrimental effect on family
relationships
 References
• Campbell, 1999, Nuclear transfer in farm animal species. Sem. Cell
Develop. Biol., 10: 245‐252.
• Gurdon, 1986, Nuclear transplantation in eggs and oocytes, J. Cell
science, Suppl. 4: 287‐318.
• •Stephens, 1995 Plant micropropagation using African Violet leaves
www.biotech.iastate.edu/publications/lab_protocols/AV_Micropropagat
ion.html
• •Vajta and Gjerris, 2006, Science and technology of farm animal
cloning: State of the art, Animal Reproduction Science, 92: 211 ‐230.
• •Wells, 2002, Nuclear transfer from established cell lines,
Encyclopedia of Life Sciences, Macmillan Publisher Group, New York,
13:327‐331.
• •Wilmut, 2003, Whole animal cloning, Encyclopedia of Life Sciences,
Macmillan Publisher Group, New York
• •Wolf et al., 1998, Nuclear transfer in mammals: Recent developments
and perspectives, J. Biotechnology, 65: 99 ‐110.
• •http://www.saskschools.ca/~stmarypa/bio30/animalcloning
THANKS