MAMMALIAN

CLONING, EMBYO
STEM CELL
TECHNOLOGIES
 REPRODUCTIVE CLONING

The technology of nuclear transfer into enucleated

oocytes (cytoplasms) was already successfully used in
1987 for reproduction of live bovine and murine
offsprings however the embryo development could
then be obtained when transplanted nucleus originated
from blastomeres,i.e cells from the first divisions
ofpreimplantation embryo development .The birth in
1997 the first sheep cloned from an adult mammary
glands cells that conveyed the full impact of the potential
of reproductive cloning since now it could be done at
any time during the life of anindividual being.
GENE/DNA CLONING
 GENE CLONING
• Gene cloning is the process in which a gene of
interest is located and copied (cloned) out of DNA
extracted from an organism. When DNA is
extracted from an organism, all of its genes are
extracted at one time. This DNA, which contains
thousands of different genes. The genetic engineer
must find the one specific gene that encodes the
specific protein of interest.
• The term genomic DNA clone or chromosomal
DNA clone then refers to an individual cell carrying
a cloning vector wit
• h one of the cellular DNA fragments.
WHAT IS GENE CLONING?
WHY WE CLONED DNA?
 THE BASICS OF CLONING
• Source of DNA: to be cloned
• Choice of vectors: to carry,maintain,and
replicate cloned gene in hpost cell
• Restriction enzyme:to cut dna
• DNA ligase:to join foreign and vector DNA
recombinant DNA
• Host cell: in which the recombinant DNA can
replicate.
 HOW IS DNA CLONED?

Cell Based DNA Cloning Cell Free DNA Cloning

 CELL BASED DNA CLONING
• DNA is extracted here from blood.
• Restriction enzyme eg:Ecor1,Hind3 etc cut the
DNA into small pieces.
• Different DNA pieces cut with the same
enzyme can join or recombine.
 CELL FREE DNA CLONING
• Developed in mid 1980’s
• Nobel prize was given to Kary Mullis in 1993
• DNA fragments can be amplified in large
amounts
• In vitro technique
 THERAPEUTIC CLONING
Cloning designed as therapy for a disease. In
therapeutic cloning, the nucleus of a cell, typically a
skin cell, is inserted into a fertilized egg whose
nucleus has been removed. The nucleated egg begins
to divide repeatedly to form a blastocyst.
How is Therapeutic Cloning
Performed?
Therapeutic cloning is another phrase for a
procedure known as somatic cell nuclear
transfer (SCNT). Here's how it works:
A scientist extracts the nucleus from an
egg
The nucleus holds the genetic material for
a human or laboratory animal
The scientist then takes a somatic cell,
which is any body cell other than an egg or
sperm, and also extract the nucleus from
this cell
 The nucleus that is extracted from the
somatic cell in the patient is then inserted
into the egg, which had its nucleus previously
removed
In a very basic sense, it's a procedure of
substitution. The egg now contains the
patient's genetic material, or
instructions
It is stimulated to divide and shortly
thereafter forms a cluster of cells known as a
blastocyst
 Benefits of Therapeutic Cloning
• Organ transplant alternative
• Leukemea
• Generative diseases
• Spinal cord repair
• Skin grafting
 Embryonic Stem
Cells For
Therapeutic Cloning
 My personal beliefs – not
necessarily yours!
• Using embryonic stem cells from aborted
fetuses is morally reprehensible to a large
enough fraction of society that it should never
become the source of choice for human stem
cells.

• Embryonic stem cells will be only a temporary,

but unfortunately probably necessary, tool
needed by researchers to learn how to modify
adult stem cells (for which there is little
controversy) for a similar use.
*But there are different types of embryonic
stem cells, some of which are ethically less
controversial than others.
 Different kinds of
stem cells
Embryonic stem cells come from embryos
(<6 weeks). Stem cells from blastocysts (2
weeks)
are virtually “immortal”.

Fetal stem cells come from fetuses (> 6

weeks)

Stem cells are present in some adult tissues,

including brain, spinal cord, and bone
marrow.
Embryonic Stem Cells
ES cells was demonstrated by formation of
ectoderm,mesoderm and endoderm
 Importance of Stem Cells
Studies of human •Human embryonic stem
embryonic stem cells cell therapies can save
will lead to major lives and restore function
advances in human of people
biology

Embryonic stem Human embryonic

cell research will stem cell can replace
provide critical damaged or lost
insights into cells
mechanisms of cell
differentiation,
growth, and death.
Understandin These include
g stem cells diabetes, degenerative
may provide neurological diseases,
keys to why demyelinative diseases,
people age. brain & spinal cord
injury.
Limitations
on the study of These conditions
human are the most
embryonic common and
stem cell costly causes of
research will disability in the
hold back United States.
biomedical
research.