SCNT and Frameshift

loning
Cloning
Clonin
SOMATIC CELL
NUCLEAR TRANSFER
(CLONING)
Overview
 What are cells? Cells are the fundamental, structural,
and functional units of living organisms.
 What is cloning? The term, “cloning,” describes several
different processes of making identical copies of
biological material.
 Why is cloning important? Because it has the potential
to treat a wide range of diseases by generating “self ”
tissues.
What Is a Cell?
 Cells are the fundamental structural, and DNA
functional units of living organisms.
 Organisms, except bacteria, are made of
cells, in which the nucleus is surrounded
Nucleus
by a membrane (eukaryotic cells). Cytoplasm
 The nucleus contains DNA, which
provides instructions for the entire
organism.
 As a cell specializes, only DNA related
to the particular functions of that cell
remains active.
Cloning Involves Making Identical Copies
“Cloning” can mean several things:
 To make many identical copies of a DNA molecule
or a particular stretch of DNA (DNA cloning or

molecular cloning).
 To replicate an entire organism (reproductive
cloning).
 To produce undifferentiated cells (stem cells) for
the purpose of studying and treating diseases

(therapeutic cloning).
Recombinant DNA Technology
Vector
(plasmid backbone)
 Recombinant DNA
technology includes DNA
+ DNA fragment cloning, gene cloning and
to be cloned molecular cloning.
 DNA from one organism is
DNA fragment ligated (inserted) transferred to a bacterial
into vector creating recombinant plasmid for replication.
DNA molecule.
 Although viruses, bacterial
To make many copies of the artificial chromosomes, and
recombinant DNA molecule, the yeast artificial chromosomes
vector (including the DNA fragment) also may be used for
is introduced into bacteria. replicating DNA, bacterial
plasmids are most
commonly used in this
Bacterial technology and are called
cell Chromosomal DNA vectors.
Brief History of Modern Cloning
 1938: Hans Spemann proposes animal cloning using somatic cell nuclear
transfer (SCNT).
 1952: Robert Briggs and Thomas King transplant a frog embryo into an
unfertilized egg without a nucleus.
 1977: Karl Illmensee claims to have cloned mice (in 1983 accused of
scientific fraud).
 1984: Davor Solter (Wistar Institute, PA) asserts cloning is biologically
impossible.
 1984: Steen Willadsen (Cambridge, UK) clones sheep embryos by
separating the cells in early embryos, similar to the way twins arise
naturally (called embryo twinning). First verified instance of a cloned
mammal.
 1986: at the University of Wisconsin, a cow is cloned from an embryo cow
cell, using embryo twinning.
 1997: Ian Wilmut (Roslin Institute) cloned a sheep, named “Dolly,” from a
mammary (breast) cell.
Somatic Cell Nuclear Transfer (SCNT)
 A somatic cell is a any cell that is not a germ cell (egg or sperm).
 Somatic Cell Nuclear Transfer (SCNT) involves:
 removing the nucleus from a somatic cell;
 inserting the nucleus into an egg cell from which the original
nucleus has been removed; and
 “shocking” the implanted cell (chemically or electrically) so it
will grow and multiply.
 The new cell is genetically identical to the donor.
 SCNT was first demonstrated in 1983 using amphibians.
 This process was attempted with mammals in 1986, but these
experiments were limited to developing embryos.
 In 1997, the first successful mammal clone from an adult somatic
cell was a sheep named Dolly. She was produced by implanting an
embryo created by SCNT into a mother sheep.
Nuclear Transfer in Progress
Somatic Cell Nuclear Transfer (SCNT):
Dolly—The First True Clone (1997)
Breast Cell (six-year-old Sheep) Get the nucleus
+
Empty
Egg Cell
Cloned Sheep!
1 29 434
Number of cloned Number of reconstructed eggs that Number of successfully
sheep actually born were able to be implanted in reconstructed eggs
pseudo-pregnant sheep
Embryonic Stem Cells Can
Become Any Tissue in the Body
Cultured Laboratory Stem Cells
Blastocyst
Scientific manipulations entice stem cells to become

specialized tissues (blood, muscle, brain etc.).
Blood Cells Muscle Cells Neuron (Brain) Cells

Stem Cells and Therapeutic Cloning
Replacing damaged cells

+ (i.e., after aggressive treatment)
Cultured laboratory
stem cells
Tissue repair
(repairing diabetic pancreas)
Tissue engineering
Blastocyst (potentially growing a new organ)
Why Use Cloning Technology?
 Using the body’s own

Cells from
patient cells to generate
Patient
tissues not only
provides a plentiful
supply but also
eliminates tissue
Therapeutic
rejection.
cloning  The technology
potentially could be
used to treat a wide
range of conditions,
from heart damage to
Patient’s cultured laboratory
stem cells used for treatment diabetes.
Therapeutic Cloning vs. Reproductive Cloning
 Not generally
Yields stem cells
condoned
that have
bythe
scientists.
potential to repair damaged,

diseased, or degenerating
Previous cell tissue.(UV light, toxic chemicals) is
damage/exposure
 Could potentially
transferred to the clone.
offer a cure for a wide range of diseases.
 No tissuefor
Potential rejection
a wideproblems.
range of health problems.
 Shortenedclinical
Practical life span
applications
(cells age more
could quickly).
take more than 10 years to

develop.
No potential for the treatment of disease. Not practical as a
 Aimed atfor
solution production
fertility problems.
of stem cells only.
 Aimed at producing a complete new individual.
Mutations: changes in DNA/RNA
caused by mistakes during
replication, transcription, or
environmental factors
Frameshift Mutations
 Nucleotide(s) are inserted or deleted

 Affects the entire amino acid
sequence, changes the reading frame
Frameshift
Insertion
A base is added and causes a change in all the amino
acids AFTER it
Frameshift
Deletion
A base is deleted and causes a change in all the amino
acids AFTER it.
Frameshift
 Frameshift mutations cause MAJOR

changes to the amino acid sequence.
*Potentially can change every amino acid in the chain
after the mutation has occurred.
THANK
YOU!!

SCNT and Frameshift

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SCNT and Frameshift

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loning

or a particular stretch of DNA (DNA cloning or

the purpose of studying and treating diseases

Breast Cell (six-year-old Sheep) Get the nucleus

Cultured Laboratory Stem Cells

Scientific manipulations entice stem cells to become

Blood Cells Muscle Cells Neuron (Brain) Cells

Replacing damaged cells

 Using the body’s own

 Nucleotide(s) are inserted or deleted

 Frameshift mutations cause MAJOR

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