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Topics of Discussion
What is cloning? Methods of cloning Dolly in detail Dollys probability Todays legality The future of cloning Ethical final questions
What is cloning?
Reproductive cloning- The entire animal is produced from a single cell by asexual reproduction. This would allow for the creation of a human being who is genetically identical to another. Therapeutic cloning- Broader use of the term cloning. Does not create a new genetically identical individual. Research includes therapy for human mitochondria disease and others that could replace damaged or diseased tissues without the risk of rejecting anothers tissue. Could create new skin tissue for burn patients.
Methods of cloning
Embryo splitting- Artificially splitting a single embryo at a very early stage of development. In the natural process this would create twins. However, because this is done at an early stage and there are usually less than eight cells you can only make a few clones. Both the nuclear genes and mitochondria genes would be identical.
Methods of cloning
Nuclear replacement- Genetic material (nucleus from embryonic, fetal, or adult cell) is removed and placed into an unfertilized egg or embryo, whose nucleus has been removed. In this case the nuclear genes remain the same but the mitochondria DNA would be different. This has the potential to create the clone of an adult organism as well as many clones at once.
Dolly in detail
Dolly was cloned using the nuclear replacement method. Again the nucleus with chromosome sets is fused with an unfertilized egg whose nucleus has been removed. Motivating factor was that it could help to improve certain qualities in livestock. Dolly was not the first sheep to be created from nuclear replacement. Two genetically identical sheep, Megan and Morag were born in 1996 using the technique. The difference was that Dolly was derived from an adult sheep, and Megan and Morag were from a sheep embryo.
How to clone?
Dollys probability
Cells taken from a six-year-old Finnish Dorset ewe and cultured in a lab. 277 cells then fused with 277 unfertilized eggs (each with the nucleus removed) 29 viable reconstructed eggs survived and were implanted in surrogate Blackface ewes. 1 gave birth to Dolly 0.361% chance at onset, 3.4482% once implanted. In nature between 33-50% of fertilized eggs develop.
Cloning Dolly
Enucleate the eggs produced by Scottish Blackface ewes (female sheep).
Treat the ewes with gonadotropin-releasing hormone ( GnRH) to cause them to produce oocytes ready to
be fertilized. Like all mammals, these are arrested at metaphase of the second meiotic division (meiosis II). Plunge a micropipette into the egg over the polar body and suck out not only the polar body but the haploid pronucleus within the egg.
Culture the cells until they have grown into a morula (solid mass of cells) or even into a blastocyst (6 days). Transfer several of these into the uterus of each (of 13, in this case) Scottish Blackface ewes (previously treated with GnRH to prepare them for implantation. Wait (with your fingers crossed). The result: one ewe gave birth (148 days later) to Dolly.
Cloning provides the most direct demonstration that all cells of an individual share a common genetic blueprint.
How do we know that Dolly is not the progeny of an unsuspected mating of the foster mother?
She has a white face and the foster mother is a Scottish Blackface DNA fingerprinting reveals bands found in Finn Dorset sheep (the breed that supplied the mammary cells), not those of Scottish Blackface sheep
The balance becomes negative as the cells age, leading to a degradation of chromosomes and to cell death after about 50 multiplications. Dolly's telomeres were short but this was also the case for the donor cell line, which was derived from an old sheep and was cultured over a long period of time.
It was prematurely suggested that cloning might generate old newborns. In all the cloned animals obtained after Dolly and in which DNA was examined, the length of telomeres was normal or longer than normal. This was also true for clones derived from a 17-yearold bull. It is also interesting to note that the two lambs born after Dolly was naturally fertilized have normal telomeres.
Highly unlikely.
In Future Attempts at human cloning are viewed very unfavorably in the scientific community.
MICROMANIPULATION
The technique or practice of manipulating cells or tissues (as by microdissection or microinjection)
Micromanipulation
INTRODUCTION Since the birth of the first baby achieved through conception outside of the human body in 1978, the principles of "in vitro" (literally "in glass") fertilization and culture have remained the same - careful establishment and maintenance of a well-controlled, sterile environment in which the normal physiology of fertilization and early development can be played out relatively undisturbed to provide healthy embryos for transfer back into the body. During the ensuing two decades, much has been learned, however, about the tolerances of such a system and how this technique can be exploited to treat a widening range of infertility cases. There have been great strides made in development of more appropriate culture media that has enabled embryos to be grown for extended periods of time in culture. Surplus embryos and possibly eggs may now routinely be cryopreserved in liquid nitrogen for use in subsequent attempts at pregnancy. Fertilization itself is no longer a hit-and-miss affair with the advent of assisted fertilization through micromanipulation. Embryos can be micro-manipulated for cell biopsy to determine their genetic status as well as aid in their ability to implant through drilling into their outer shell (assisted hatching).
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