History of Human Cloning
History of Human Cloning
History of Human Cloning
Sea urchin
The sea urchin is a relatively simple organism that is useful for studying
development. Dreisch showed that by merely shaking two-celled sea urchin
embryos, it was possible to separate the cells. Once separated, each cell grew
into a complete sea urchin.
This experiment showed that each cell in the early embryo has its own
complete set of genetic instructions and can grow into a full organism.
Salamander
Hans Spemann
Spemann’s first challenge was to figure out how to split the two cells of an
embryo much stickier than sea urchin cells. Spemann fashioned a tiny noose
from a strand of baby hair and tightened it between two cells of a salamander
embryo until they separated. Each cell grew into an adult salamander.
Spemann also tried to divide more advanced salamander embryos using this
method, but he found that cells from these embryos weren’t as successful at
developing into adult salamanders.
This experiment showed that embryos from a more-complex animal can also
be “twinned” to form multiple identical organisms—but only up to a certain
stage in development.
Hans Spemann
Again using a strand of baby hair tied into a noose, Spemann temporarily
squeezed a fertilized salamander egg to push the nucleus to one side of the
cytoplasm. The egg divided into cells—but only on the side with the nucleus.
After four cell divisions, which made 16 cells, Spemann loosened the noose,
letting the nucleus from one of the cells slide back into the non-dividing side of
the egg. He used the noose to separate this “new” cell from the rest of the
embryo. The single cell grew into a new salamander embryo, as did the
remaining cells that were separated.Essentially the first instance of nuclear
transfer, this experiment showed that the nucleus from an early embryonic cell
directs the complete growth of a salamander, effectively substituting for the
nucleus in a fertilized egg.
Briggs and King transferred the nucleus from an early tadpole embryo into an
enucleated frog egg (a frog egg from which the nucleus had been removed).
The resulting cell developed into a tadpole.
The scientists created many normal tadpole clones using nuclei from early
embryos. But just like Spemann’s salamander experiments, cloning was less
successful with donor nuclei from more advanced embryos: the few tadpole
clones that did survive grew abnormally.
Most importantly, this experiment showed that nuclear transfer was a viable
cloning technique. It also reinforced two earlier observations. First, the nucleus
directs cell growth and, ultimately, an organism’s development. Second,
embryonic cells early in development are better for cloning than cells at later
stages.
John Gurdon
This experiment showed that, despite previous failures, nuclei from somatic
cells in a fully developed animal could be used for cloning. Importantly, it
suggested that cells retain all of their genetic material even as they divide and
differentiate (although some wondered if the donor DNA came from a stem
cell, which can differentiate into multiple types of cells).
Rabbit
J. Derek Bromhall
Mammalian egg cells are much smaller than those of frogs or salamanders, so
they are harder to manipulate. Using a glass pipette as a tiny straw, Bromhall
transferred the nucleus from a rabbit embryo cell into an enucleated rabbit
egg cell. He considered the procedure a success when a morula, or advanced
embryo, developed after a couple of days.This experiment showed that
mammalian embryos could be created by nuclear transfer. To show that the
embryos could continue developing, Bromhall would have had to place them
into a mother rabbit's womb. He never did this experiment.
Sheep
Steen Willadsen
Willadsen used a chemical process to separated one cell from an 8-cell lamb
embryo. The he used a small electrical shock to fuse it to an enucleated egg
cell. As luck would have it, the new cell started dividing.
By this time, in vitro fertilization techniques had been developed, and they had
been used successfully to help couples have babies. So after a few days,
Willadsen placed the lamb embryos into the womb of surrogate mother sheep.
The result was the birth of three live lambs.
Cow
Using methods very similar to those used by Willadsen on sheep, First, Prather,
and Eyestone produced two cloned calves. Their names were Fusion and Copy.
This experiment added cows to the list of mammals that could be cloned by
nuclear transfer. Still, mammalian cloning was limited to using embryonic cells
as nuclear donors. Cloning using nuclei from differentiated adult somatic cells
still wasn’t thought possible.
Sheep
All previous cloning experiments used donor nuclei from cells in early embryos.
In this experiment, the donor nuclei came from a slightly different source:
cultured sheep cells, which were kept alive in the laboratory.
Wilmut and Campbell transferred the nuclei from cultured cells into
enucleated sheep egg cells. The lambs born from this procedure were named
Megan and Morag.
This experiment showed that cultured cells can supply donor nuclei for cloning
by nuclear transfer. Because scientists had already learned how to transfer
genes into cultured cells, this experiment showed that it might be possible to
use such modified cells to create transgenic animals—such as cows that could
make insulin for diabetics in their milk.
Sheep
Of 277 attempts, only one produced an embryo that was carried to term in a
surrogate mother. This famous lamb, named Dolly, brought cloning into the
limelight. Her arrival started conversations about the implications of cloning,
bringing controversies over human cloning and stem cell research into the
public eye.
After developing a lung disease called jaagsiekte, she was euthanized on Feb.
14, 2003, stuffed, and put on display at the Museum of Scotland in Edinburgh,
where you can find her in a new science gallery starting on Friday.
http://www.businessinsider.com/can-you-clone-a-human-2016-7/?IR=T
11.1997 - First primate created by embryonic cell nuclear transfer
Rhesus monkey
Primates are good models for studying human disorders. Cloning identical
primates would decrease the genetic variation of research animals, and
therefore the number of animals need in research studies.
Sheep
To create the transgenic sheep, the scientists performed nuclear transfer using
donor DNA from the cultured transgenic cells. The result was Polly, a sheep
that produced Factor IX protein in her milk.
This experiment showed that sheep could be engineered to make therapeutic
and other useful proteins in their milk, highlighting the potential medical and
commercial uses for cloning.
Multiple groups
After the successes leading up to Dolly and Polly, other scientists wanted to see
if similar techniques could be used to clone other mammalian species. Before
long, several more animals had been successfully cloned. Among them were
transgenic animals, clones made from fetal and adult cells, and a male mouse;
all previous clones had been female.
Multiple groups
Rhesus monkey
Shoukhrat Mitalipov and colleagues
Researchers took a cell from an adult monkey and fused it with an enucleated
egg cell. The embryo was allowed to develop for a time, then its cells were
grown in a culture dish. These cells, because they can differentiate to form any
cell type, are called embryonic stem cells.
Human
In this experiment, researchers took a skin cell from the patient and fused it
with a donated egg cell. Key to the success of the experiment were
modifications to the culture liquid in which the procedure was done and to the
series of electrical pulses used to stimulate the egg to begin dividing.
http://learn.genetics.utah.edu/content/cloning/clonezone/
what is cloned? Clone – in biology, a clone is defined as an organism
having identical or nearly identical genetic material. Therefore, clones
are not only the organisms obtained in the laboratory as a result of
complex procedures, but also organisms created in the process of
vegetative reproduction, such as bacteria, unicellular organisms,
offshoots and plantlets.
Cloning – the procedure for obtaining organisms with the same genetic
information. How is it done? You need to collect an egg cell from a
donor (a female sheep, mouse or cat). Then you need to carefully
remove the nucleus from the cell and collect another cell from the skin,
udder or other tissue from another male or female donor of the same
species (i.e. from another sheep, mouse or cat). This is the animal that
will be cloned. From this cell, you also need to remove the cell nucleus
and place it in the empty egg. The egg obtained in such a way needs to
be treated with a gentle electric shock. The egg should begin to divide
and grow into a multicellular embryo. At this stage, the embryo needs to
be implanted into the uterus of a surrogate mother. If the pregnancy
develops normally and the animal is born – we have a clone!
http://www.kopernik.org.pl/en/special-projects/archiwum-
projektow/projekt-genesis/krotka-historia-klonowania/
The first human cloning
The birth of the world’s first cloned human was condemned by British
scientists today as another example of the “sordid depths” to which maverick
physicians will sink. The claim by Clonaid announcing Boxing Day’s arrival of a
baby girl called Eve born by caesarean section, will prompt “revulsion and
disgust” throughout the world and will see 2003 go down in history of mankind
as the Year of the Clone, Dr Patrick Dixon claimed.
“We don't know yet (27/12/02) whether the claim about Eve is fact or fraud
but one thing is clear. There’s a global race by maverick scientists to produce
clones, motivated by fame, money and warped and twisted beliefs,” he said.
“Today’s announcement is totally inevitable and we can expect a number of
other births of clones to be announced over the next few weeks.”
He said that physicians across the world were propelled by “private passions
and weird emotions” with the determination to deliver a cloned baby to any
man or woman who wished to “duplicate themselves or recover the dead”.
The cloning industry, and today’s announcement, was worth tens of millions of
pounds, he said.
The Raelian Sect, who claimed responsability for the first human clone, insist
that an independent inspector will be employed to take DNA evidence from
the baby and her 30-year-old mother. (Editor: they then went back on that
promise 7 days later).
Dr Dixon added: “The baby born has been born into a living nightmare with a
high risk of malformations, ill-health, early death and unimaginable severe
emotional pressures. We should be very concerned for Eve 's welfare.
“Can you imagine what it will be like for a 12-year-old daughter to look at her
mother and realise she is seeing her own sister? “For Eve to look at her own
grandparents around the Christmas table and to see her parents? Can you
imagine what kind of freak show she will be regarded as for her entire lifetime?
“What will it do to her sense of personal identity, knowing that she’s only a
copy of someone else who is much older?”
“They are trying to recover the dead. One person told me how his 18-year-old
handicapped son and died and he wanted to know what he would have been
like had he not been starved of oxygen at birth.
“The only reasons people want to clone are for selfish reasons. They are totally
obsessed with their own right to have a clone or to clone dead realtives,
without any regard to the welfare of the child, and this is the whole thrust of
it.”
Dr Dixon claimed that although the baby had not been born in Britain, if the
claims turned out to be genuine, it was probably British technology which had
guided the process.
“The technology is well proven, not only in animals but also in humans. Last
year Advanced Cell Technology (ACT) in the US announced they had
successfully made the world's first cloned embryos using human eggs. Indeed,
the reason why UK parliament gave a green light to proceed with legal creation
of cloned embryos was because they had been told by British cloners that it
was possible and important to do it for medical research.
(Note: Chinese scientists announced end January 2003 that they had
cloned 80 human embryos of which 4 developed far enough to be
implanted, before being destroyed. )
Australian government gives first license to create cloned human
embryos to try and obtain embryonic stem cells. (17 Sept 2008)
The big question is what has been going on over the last two or three years? If
human clones have already been made, how would you know?
People imagine you can clone tissues for treatment without making whole
embryos but you can't. The technology is identical - whether you implant a
clone to be born, cull it for spares or cannibalise it before implantation to make
a human tissue factory out of embryonic stem cells.
You can't grow organs from cells. If you want organs you have to take them
from a late pregnancy foetus. You will only grow, perhaps, bone marrow or
nerve tissue.
Unwanted, but no one knows what to do. Human embryos are more than bags
of biodata, wherever you stand on pro-choice or pro-life. They have all the
promise of a beautiful baby son or daughter. There is a profound mystery here:
human life developing from a single cell.
As a doctor I know we need gene technology to feed the world and cure
disease but we don't need human cloning. We need a biotech summit, a global
ban to strongly discourage cloners from making babies and a halt on further
research until debate is concluded and laws are in place. Over 170 nations
have no gene laws. Regulation in America is almost useless when you can hop
on a plane and implant a clone in an hour elsewhere. (Dr Patrick Dixon, Los
Angeles Times December 1998)
https://www.globalchange.com/clonelatimes.htm
Tahap I – Tahap awal adalah denaturasi DNA yang artinya DNA harus
ditingkatkan terlebih dahulu suhunya yang kurang lebih 94 derajat Celcius
dalam jangka waktu 1 menit agar terjadi pemisahan antara 2 DNA dupleks.
Tahap III – Tahap ketiga adalah pemanjangan rantai DNA dengan cara
memanaskannya kembali pada suhu 72 derajat Celcius dalam waktu 1 menit.
Perlu diingat bahwa pada tahap ini setidaknya diulang sebanyak 25-35 kali.
Dapat dijabarkan lebih lanjut bahwa pada setiap DNA dupleks yang yang
terdapat pada tahap pertama tersebut masing masing akan membentuk 2
untai anak DNA, sehingga menghasilkan 4 untai DNA. Lalu pada tahapan
kedua, 4 untai DNA tersebut akan menghasilkan 8 untai DNA. Dan pada tahap
akhir dari tahap kedua nantinya akan terbentuk polinukleotida dengan panjang
yang sangat terbatas antara 1 DNA primer dengan yang lainnya. Fragmen
fragmen DNA ini kemudian disebut sebagai amplikon. Fragmen DNA akan
digandakan lebih banyak lagi dan nantinya akan menghasilkan DNA yang
sangat lengkap yang merupakan hasil dari DNA sampel diawal.
Proses kloning pada manusia tahap kedua adalah penyisipan fragmen DNA
atau ligasi merupakan tahapan penggabungan antara fragmen DNA satu
dengan yang lainnya sehingga nantinya akan tercipta DNA rekombinan. Secara
umum, ligasi framen DNA dengan cara in vitro ini memiliki 3 cara, yaitu:
Cara I – Cara pertama adalah dengan menggunakan enzim DNA ligase yang
berasal dari bakteri. ( baca : Reproduksi Bakteri )
Cara II – Cara kedua adalah dengan menggunakan DNA ligase yang berasal dari
bakteri E-Coli yang sudah terinfeksi oleh bakteriofag T4 atau biasa disebut
enzim T4 ligase. ( baca : Peran Bakteri yang Menguntungkan )
Cara III – Cara ketiga adalah dengan cara memberi enzim deoksinukleotidil
transferase agar fragmen DNA tersebut dapat tersintesis dengan baik.
Yang perlu diingat disini adalah bahwa pada cara pertama hanya digunakan
untuk meligase ujung DNA yang lengket, sedangkan pada cara kedua dapat
digunakan pada DNA yang memiliki ujung lengket ataupun tumpul. Disisi lain,
pada cara ketiga digunakan untuk mensintesis untai DNA tunggal
homopolimerik 3′. Adapun hal lain yang perlu diperhatikan betul adalah
tentang suhu optimum DNA.
Suhu optimum dari DNA ligase akan berkisar pada suhu 37 derajat Celcius,
dimana pada suhu tersebut biasanya hidrogen akan membentuk suatu ikatan
dengan sendirinya yang dapat ditemukan di bagian ujung lengket DNA, akan
tetapi hal ini membuatnya menjadi tidak stabil dan dapat menyebabkan
kerusakan pada DNA karena suhu yang terlalu tinggi ditempat ikatan tersebut.
Maka dari itu, proses ligase ini biasanya akan berlangsung pada suhu 4-15
derajat Celcius dalam jangka waktu 24 jam.
Agar ligasi dapat bekerja secara efisien, maka dapat menggunakan fragmentasi
DNA yang memiliki konsentrasi tinggi yaitu lebih dari 100µg/ml atau dengan
menggunakan enzim alkalin fosfatase dengan tujuan untuk menghilangkan
gugus fosfat yang dimulai dari ujung 5′ yang berada pada molekul DNA.
Selanjutnya dapat menggunakan cara penambahan molekul linker, molekul
adaptor demi mensintesis untai tunggal homopolimerik 3′. Dengan demikian
DNA rekombinasi dapat terbentuk dengan susunan 5’ GAATTC 3’ — 3’CTTAAG
5′.
3. Transformasi DNA
Sedangkan pada transformasi buatan, sel bakteri akan diubah menjadi sel yang
kompeten dan akan masuk ke selubung sel bakteri dengan sifat yang
permeabel, kemungkinan terbesarnya adalah DNA tersebut akan mengikat
dengan sel lalu masuk ke bagian sitoplasma dan bereaksi dengan sel genom
bakteri. Yang perlu diketahui disini adalah bahwa proses memasukkan sel
kompeten tersebut dilakukan dengan alat kejut. ( baca : Fungsi Membran Sel )
Adapun tahap transformasi DNA yang secara umum adalah sebagai berikut:
Pada mulanya akan terjadi pengenalan serta penempelan dsDNA dari luar ke
dinding sel bakteri ( baca : Struktur Tubuh Bakteri )
dsDNA tersebut kemudian masuk ke dinding dan membran sel melalui channel
protein
dsDNA nantinya akan mengalami degradasi di salah satu bagian untainya dan
menyisakan ssDNA yang nantinya akan masuk ke sitoplasma
Dalam sitoplasma inilah nantinya ssDNA akan menjadi template yang akan
digunakan dalam proses sintesis dsDNA yang baru dimana nantinya akan
terintegrasi dalam satu kromosom serta menjadi plasmid yang baru ( baca :
Fungsi Kromatin )
4. Seleksi Kloning
Tahap terakhir dari proses kloning pada manusia adalah penyeleksian hasil
kloning. Tujuan dari seleksi ini adalah agar koloni bakteri yang tidak diinginkan
dapat dipotong dengan bantuan enzim retriksi. Penyeleksian tersebut biasanya
menggunakan sistem X-gal untuk mengidentifikasi bagian plasmid yang
rekombinan dengan komplementasinya. Hasil pemotongan tersebut nantinya
akan dielektroforesis agar pita fragmen DNA dapat terlihat dan dapat
dipisahkan.
Sel Stem – Pada mulanya siapkan sel stem yang merupakan sel yang nantinya
akan tumbuh menjadi berbagai bagian sel tubuh terlebih dahulu.
Pengambilan Inti Sel – Setelah sel stem sudah siap, maka dilakukan
pengambilan inti sel yang didalamnya terdapat info genetik. ( baca : Kelainan
Kromosom )
Sel Telur – Sementara inti sel stem dipisahkan, siapkanlah sel telur yang akan
digunakan dan intinya juga harus dipisahkan.
Perangsangan Sel Telur – Setelah kedua inti sel tersebut bertemu, maka sel
telur harus dipicu agar dapat melakukan pembelahan dan akan menjadi
embrio. ( baca : Bagian Bagian Membran Embrio )
https://dosenbiologi.com/bioteknologi/proses-kloning-pada-manusia
Dampak positif :
2. Dampak negatif :
6. Individu hasil kloning sel-selnya diperoleh dari induknya. Ini berarti umur
sel-sel hasil kloning pun sama dengan umur sel-sel induknya. Oleh karena itu,
individu hasil kloning pun akan memiliki umur sama dengan induknya.
http://borgomaulyn.blogspot.co.id/2014/03/dampak-positif-dan-negatif-
kloning.html