WHAT IS CLONING?

Cloning is the asexual production of an exact copy of an original. So for example, one could use
cloning to produce the exact copy of a single cell. The cell copy would be identical to the first
cell and would have the same exact DNA sequence. In many cases, cloning has been used to
reproduce type specific cells. In some instances, cloning of an individual organism.

Unlike reproduction that involves two “parents,” such as a male and female plants, cloning has a
single parent. This is often used in reproducing certain plants. Certain plants have undergone
cloning processes for thousands of years, but they do not play a part in the ethical debates that
surround cloning of animals, and most particularly humans.

For example, reproductive cloning of animals was first attempted in the 1950s. Most identify the
sheep Dolly, cloned in 1996. Dolly’s parent had DNA transferred into an egg that had its nucleus
removed. This is called a somatic cell nuclear transfer. The cell was then treated with chemicals
and stimulated to grow so than an almost exact replicate of the cloned sheep was born.

However, the idea of cloning humans still remains. While many people feel that cloning human
tissue, as for organs for transplant might be valuable, many others feel that cloning a whole
human is unethical. Some scientists without religious affiliation also believe that ethical issues
that might be engendered in prolonging life through cloned tissues need further scrutiny.

Some further feel that the harvesting of stem cells from an embryo is also wrong, or that
creating embryos for the purpose of harvesting stem cells is unethical. Others argue that stem
cell research may point the way toward curing diseases for which there is currently no cure. It
should be noted, however, that fewer people object to the idea of cloning a body part, than
cloning a human.

In some countries, stem cell research has been halted, when it involves cloning human
embryos. Other scientists investigate the possibility of finding stem cells elsewhere, as in the
umbilical cord blood of newborns. It is suspected that some countries may be attempting to
clone a whole human, but have not yet achieved this.
HOW IS ANIMAL CLONING CARRIED
OUT?

You may have first heard of cloning when Dolly the Sheep showed up on the scene in 1997.
Cloning technologies have been around for much longer than Dolly, though.

How does one go about making an exact genetic copy of an organism? There are a couple of
ways to do this: artificial embryo twinning and somatic cell nuclear transfer. How do these
processes differ?

1. Artificial Embryo Twinning

Artificial embryo twinning is the relatively low-tech version of cloning. As the name suggests,
this technology mimics the natural process of creating identical twins.

In nature, twins occur just after fertilization of an egg cell by a sperm cell. In rare cases, when
the resulting fertilized egg, called a zygote, tries to divide into a two-celled embryo, the two cells
separate. Each cell continues dividing on its own, ultimately developing into a separate
individual within the mother. Since the two cells came from the same zygote, the resulting
individuals are genetically identical.

Artificial embryo twinning uses the same approach, but it occurs in a Petri dish instead of in the
mother's body. This is accomplished by manually separating a very early embryo into individual
cells, and then allowing each cell to divide and develop on its own. The resulting embryos are
placed into a surrogate mother, where they are carried to term and delivered. Again, since all
the embryos came from the same zygote, they are genetically identical.

2. Somatic Cell Nuclear Transfer

Somatic cell nuclear transfer, (SCNT) uses a different approach than artificial embryo twinning,
but it produces the same result: an exact clone, or genetic copy, of an individual. This was the
method used to create Dolly the Sheep.

However, what is somatic cell nuclear transfer ?

Somatic cell: A somatic cell is any cell in the body other than the two types of reproductive
cells, sperm and egg. Sperm and egg are also called germ cells. In mammals, every somatic
cell has two complete sets of chromosomes, whereas the germ cells only have one complete
set.

Nuclear: The nucleus is like the cell's brain. It's an enclosed compartment that contains all the
information that cells need to form an organism. This information comes in the form of DNA. It's
the differences in our DNA that make each of us unique.
Transfer: Moving an object from one place to another.

To make Dolly, researchers isolated a somatic cell from an adult female sheep. Next, they
transferred the nucleus from that cell to an egg cell from which the nucleus had been
removed. After a couple of chemical tweaks, the egg cell, with its new nucleus, was behaving
just like a freshly fertilized zygote. It developed into an embryo, which was implanted into a
surrogate mother and carried to term.

The lamb, Dolly, was an exact genetic replica of the adult female sheep that donated the
somatic cell nucleus to the egg. She was the first-ever mammal to be cloned from an adult
somatic cell

How does SCNT differ from the natural way of making an embryo?

The fertilization of an egg by a sperm and the SCNT cloning method both result in the same
thing: a dividing ball of cells, called an embryo. So what exactly is the difference between these
methods?

An embryo is composed of cells that contain two complete sets of chromosomes. The difference
between fertilization and SCNT lies in where those two sets originated.

In fertilization, the sperm and egg both contain one set of chromosomes. When the sperm and
egg join, the resulting zygote ends up with two sets - one from the father (sperm) and one from
the mother (egg).

In SCNT, the egg cell's single set of chromosomes is removed. It is replaced by the nucleus
from a somatic cell, which already contains two complete sets of chromosomes. Therefore, in
the resulting embryo, both sets of chromosomes come from the somatic cell.
WHAT IS TISSUE CULTURE
TECHNIQUE ?

Tissue culture (often called micropropagation) is a special type of asexual propagation where
a very small piece of tissue (shoot apex, leaf section, or even an individual cell) is excised (cut-
out) and placed in sterile (aseptic) culture in a test tube, petri dish or tissue culture container
containing a special culture medium.

 The culture medium contains a gel (agar) with the proper mixture of nutrients, sugars,
vitamins and hormones, which causes the plant part to grow at very rapid rates to produce new
plantlets.  It has been estimated that one chrysanthemum apex placed in tissue culture could
produce up to 1,000,000 new plantlets in one year.  Thus, tissue culture is used for rapid
multiplication of plants.  A very specialized laboratory is required for tissue culture.  All the
procedures are done in a laboratory and special ventilated cabinet that is as sterile as an
operating room.
HOW IS CULTURE TEHCNIQUE IS
CARRIED OUT ?

Explant:  Cut-out Plant Tissue and Place in  Tissue Culture Container
The first step is to obtain what is called and explant.  This means to simply cut-out a very small
piece of leaf or stem tissue, or even isolate individual cells, and place them in a tissue culture
container.  The tissue has to be sterilized so it will not have any contaminating bacteria or
fungus.  It is then placed inside the tissue culture contain on a gel called agar.  In the agar is
dissolved all the sugar, nutrients and hormones the plant needs.

Multiplication:  Tissue Grows and Produces Small Plants

The tissue will begin to grow.  It may make a big blob of tissue called callus, or it may make new
shoots directly from the explant tissue that was inserted in the container.

Rapid Multiplication by Transfer of Cultures

Once the plantlets start developing, some can be removed and placed in new tissue culture
containers.  Thus, another "forest"' of plants is produced.  This results in a rapid multiplication of
the cultures and many thousand of plants can be produced in a few months.

Transplanting
When the plantlets are large enough, they can be removed from the tissue culture container and
transferred into pots with potting soil.  The young plants are growth in a greenhouse just like you
would any young seedling or cutting.

When the small plant clones are removed from the culture containers, they must be
transplanted into some type of acclimation container or  kept under a mist system until the
acclimate to the ambient environment.

After acclimation, the young plants can be transplanted

and grown in pots in a greenhouse to produce new plants.
WHAT IS GENETIC ENGINEERING CULTURE ?

We find it mixed in our food on the shelves in the supermarket--genetically engineered soybeans
and maize. We find it growing in a plot down the lane, test field release sites with genetically
engineered rape seed, sugar beet, wheat, potato, strawberries and more. There has been no
warning and no consultation.

It is variously known as genetic engineering, genetic modification or genetic manipulation. All

three terms mean the same thing, the reshuffling of genes usually from one species to another;
existing examples include: from fish to tomato. Genetic engineering (GE) comes under the broad
heading of biotechnology.

Genetic engineering (GE) is used to take genes and segments of DNA from one species, e.g. fish,
and put them into another species, e.g. tomato. To do so, GE provides a set of techniques to cut
DNA either randomly or at a number of specific sites. Once isolated one can study the different
segments of DNA, multiply them up and splice them (stick them) next to any other DNA of
another cell or organism. GE makes it possible to break through the species barrier and to shuffle
information between completely unrelated species; for example, to splice the anti-freeze gene
from flounder into tomatoes or strawberries, an insect-killing toxin gene from bacteria into maize
or cotton or rape seed.

Yet there is a problem - a fish gene will not work in tomato unless I give it a promoter with a
"flag" the tomato cells will recognise. Such a control sequence should either be a tomato
sequence or something similar. Most companies and scientists do a shortcut here and don't even
bother to look for an appropriate tomato promoter as it would take years to understand how the
cell's internal communication and regulation works. In order to avoid long testing and adjusting,
most genetic engineering of plants is done with viral promoters. Viruses - as you will be aware -
are very active. Nothing, or almost nothing, will stop them once they have found a new victim or
rather host. They integrate their genetic information into the DNA of a host cell (such as one of
your own), multiply, infect the next cells and multiply. This is possible because viruses have
evolved very powerful promoters which command the host cell to constantly read the viral genes
and produce viral proteins. Simply by taking a control element (promoter) from a plant virus and
sticking it in front of the information block of the fish gene, you can get this combined virus/fish
gene (known as a "construct') to work wherever and whenever you want in a plant.

This might sound great, the drawback though is that it can't be stopped either, it can't be switched
off. The plant no longer has a say in the expression of the new gene, even when the constant
involuntary production of the "new" product is weakening the plant's defences or growth.

And furthermore, the theory doesn't hold up with reality. Often, for no apparent reason, the new
gene only works for a limited amount of time and then "falls silent". But there is no way to know
in advance if this will happen.
Though often hailed as a precise method, the final stage of placing the new gene into a receiving
higher organism is rather crude, seriously lacking both precision and predictability. The "new"
gene can end up anywhere, next to any
gene or even within another gene, disturbing its function or regulation. If the "new" gene gets
into the "quiet" non-expressed areas of the cell's DNA, it is likely to interfere with the regulation
of gene expression of the whole region. It could potentially cause genes in the "quiet" DNA to
become active.

Often genetic engineering will not only use the information of one gene and put it behind the
promoter of another gene, but will also take bits and pieces from other genes and other species.
Although this is aimed to benefit the expression and function of the "new" gene it also causes
more interference and enhances the risks of unpredictable effects.