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Introduction to Cloning. What is a Clone? Colloquial definition: a copy of an individual Biological definitions: Asexual duplicate of an organism or cell; typically through mitosis/ Clonal expansion Parthenogenetic duplication of an entire organism Monozygotic twins/ Embryo splitting
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What is a Clone? Colloquial definition: a copy of an individual Biological definitions: • Asexual duplicate of an organism or cell; typically through mitosis/ Clonal expansion • Parthenogenetic duplication of an entire organism • Monozygotic twins/ Embryo splitting • Molecular cloning and subcloning • Vegetative growth
Why Clone? Reproductive cloning • Asexual reproduction of an entire animal from a single cell's genetic information. • Allowed to come to term • For infertility or resurrection of endangered species. • For in vitro manipulation of livestock • For selection of traits of livestock • Study imprinting • Study cancer • Study embryology and development/ cellular genomics Therapeutic cloning • Broad use of the word “cloning”. • Not used to create an entirely new individual. (Never implanted) • Used to create stem cells for tissue engineering to treat anything from neurodegenerative disease to organ transplantation without rejection. • Gene therapy
Parthenogenesis “Virgin Birth” • New Mexico Whiptail is all female species • Derived from female Western and male Striped Whiptail • Unfertilized eggs yield clones Pseudocopulation no exchange of genetic material progesterone estrogen Unfertilized eggs Clones of the mother
Early attempts at vertebrate cloning by Hans Spemann (1920s) By John Gurdon (1960s) Embryo Splitting Somatic Cell Nuclear Transfer UV irradiate nucleus Frog blastula Intestine, heart, skin cells Injection to enucleated oocytes Injection into enucleated oocytes Small percentage survive
Adult Somatic Cell Nuclear Transfer Ablation of nucleus from polar body Morula or Blastocyst implanted into Uterus G0 udder cells Cell fusion and activation
Dolly • Cells derived from a 6 year old Finnish Dorset ewe (white faced) • Implanted into Black-face surrogates • 277 cell fusions with 29 viable embryos • Dolly developed arthritis and lung cancer at the age of 6 • Finnish Dorset ewes live 11-12 years • Dolly was euthanized at half her breed's expected life-span
Proliferation of cloned mammals mouse, cow, gaur, mule, cat Noah, the gaur • First attempt to revive endangered species • Cloned from a gaur skin nucleus • Into enucleated cow oocyte • Died 48 hours after birth from respiratory issues Cc, the tabby cat • Cloned from Rainbow, a calico cat • Superficially nonidentical • Genotyping reveals genetic identity with Rainbow Cloned by Genetic Savings & Clone Cloned by Advanced Cell Technology
Cloning problems • Expensive • Low yield/ success rate • Shortened life-span • Cloned calves die young and are often large • Abnormal gene expression found in cloned mice • Sperm imprinting cannot be studied
Chromatin Transfer Detergent permeabilizes membrane Chromosome Condensation Impermeable cell Fusion to enucleated oocyte Cloned by Genetic Savings & Clone
Human Cloning Non implanted somatic cell nuclear transfer clone for stem cells in South Korea. Limited therapeutic cloning licenses to establish stem cells in UK. Cloning from disease patients (diabetes, Parkinson's, etc.) being sought out at Harvard
From Rael.org and Clonaid.com The Raelian Revolution, the world's largest Atheist, non-profit UFO related organisation - over 60,000 members in 90 countries - working towards the first embassy to welcome people from space... destroying the myth of god and sweeping the world with the most politically incorrect and fearlessly individualistic philosophy of non-conformism. CLONAID™ ,the first human cloning company in the world, was founded in February 1997, by RAËL and a group of investors who created the Valiant Venture Ltd Corporation based in the Bahamas. Thousands of families have contacted Clonaid in the last couple of years in the hope to see a later born twin of their deceased family member be born again. Many of those families are families of celebrities, business leaders and political leaders - we are currently working on the case of a prime minister whose son has been killed. Since March 1st, 30 human clones have been implanted, 23 pregnancies are under way. 13 cloned babies are now alive thanks to our dedicated team of doctors and scientists. They are all healthy and no problem has been found in any one of them that could be attributed to their mode of conception. The 8 babies born in February and March of this year are located in Australia, Mexico ( 2 of them), Brazil, Spain, Italy, England and Hong Kong,(sic)