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AS 91607 (3.7) Demonstrate Understanding of Human Manipulations of Genetic Transfer and its Biological Implications Internally Assessed, 3 credits Reference Text – ESA Study Guide, chapter 24 Whole Organism Cloning o Refers to the production of genetically identical whole organisms (clones). o It involves growing a whole organism from one cell taken from the organism (“parent”) that is being cloned. o Cloning is designed to quickly and cost effectively get many identical copies of an organism with desirable traits, especially transgenic organisms. Biological Implication The resulting cloned population lacks genetic diversity compared with other populations. Biological Implication Gene expression can be influenced by factors in the internal environment of the animal (epigenetics) Eg the amount of protein or trace elements in the diet. A cloned animal may not be identical to the original parent. Eg clones typically vary in colour patterns compared with the parent and with one another. Cloning may also be used in conservation. Eg cloning the surviving members of an endangered species to increase the population reduces the chances of extinction. In NZ, the last members of the Enderby Island breed of cattle (a cow “Lady”) was cloned in 1998 (producing a daughter “Elsie). It has even been proposed that cloning could be used to bring extinct species, such as woolly mammoths “back to life” (“de-extinction”) Reading – Enderby Island Cattle While Elsie is genetically identical to Lady, her colour patterns are similar but not identical. A son, “Derby” had also been born to Lady, as a result of embryo transplantation using semen recovered from Enderby Island during the culling of the cattle (culling was carried out in the early 1990’s for conservation purposes. Cloning has also been proposed for a beloved pet (either before or after death), the cloning of famous animals has such as the long dead NZ racehorse “Phar Lap” (“Big Red”) Phar Lap @ Te Papa Museum Animal Cloning Cloning whole vertebrate animals from adult donor cells is a more complex process. Because adult animal cells are specialised in that only genes related to that cell’s functions are normally expressed. However, the creation of Dolly, the cloned sheep, in 1997 from the udder cells of a Finn-Dorset ewe showed that it is possible to reprogramme an adult cell to generate an entire organism, and clones of many animals including sheep, mice, rhesus monkey, cattle, dogs, goat, camels and wolves have been produced over the past two decades. The usual method for cloning animals involves somatic cell nuclear transfer (SCNT). Dolly was a female domestic sheep, and the first mammal to be cloned from an adult somatic cell, using the process of nuclear transfer. Wikipedia Born: July 5, 1996, Scotland, United Kingdom Died: February 14, 2003 Species: Ovis aries Children: Bonnie, Rosie, Lucy, Sally, Darcy, Cotton SCNT video SCNT Nucleus from the donor adult somatic cell is transferred into an enucleated egg cell (nucleus has been removed or destroyed by UV light). A pulse of electricity fuses the two cells together (“fertilisation”) and the resultant egg is allowed to divide under artificial conditions for around 5 days. This is now known as the blastocyst. The blastocyst is then implanted into the uterus of a surrogate female for development, growth and repair. Biological Implications – Whole Organism Cloning Biological implications of cloning include possible impacts on: Genetic diversity Ecosystem Evolution Health & survival of individuals Health & survival of populations Impact on Genetic Diversity Cloning produces individuals that are genetically identical (including sex). A cloned population is less likely to survive on environmental change, especially the introduction of a new pathogen. Any species brought back to life may not walk the Earth again for any length of time – there could be a series of extinctions ad re-extinctions. Impact on Evolution Any population derived from a clone will have little or no genetic diversity. This greatly increases the risk of “reextinction” Impact on Health If the genome of the cloned individual contains contains alleles for genetic disorders, the clones inherit this disorder. Impact on Ecosystem Monocultures are plants that very similar, or identical genetically. A large population of cloned plants could be considered an extreme example of monoculture. Researchers would need to consider the following when analysing the possible impact on ecosystem from de-extinction: Will the cloned species be able to reoccupy its original niche within the biological community? Or adapt to a new niche? Is the original still available? Will the cloned species out-compete existing members of the community, and/or disrupt food chains and/or cause local extinctions? Will the cloned species be out-competed by the existing species? Will food supplies still be in existence and sufficient for the cloned species to survive? Will clones be dependent on human management for their survival, that’s is, have to occupy a managed ecosystem rather than a natural ecosystem? When it coms to cloning extinct organisms, we definitely need to proceed with caution. Another method that has been used since the 1980’s to produce animal clones is that of embryo splitting or embryo twinning. Embryos at a very early stage of development, say around 8 cell stage are still totipotent – are split to produce identical twin embryos that are then inserted into a female animal for development. Reference http://animalscience.ucdavis.edu/animalbiotech/images/Image5.gif De-extinction “Bringing them back to Life” Each year an estimated 10 000 to 100 000 animal species die off, joining the countless species that have gone extinct throughout the Earth’s history!!! Extinction is forever………..maybe not anymore!!!! Scientists are close to bringing extinct species back to life “de-extinction”. This involves finding & extracting DNA from preserved tissue remains (usually from being frozen). A single cell or intact nucleus may be sufficient, and sequencing the DNA to obtain a complete (or sufficiently complete) genome. This may involve amplification using PCR technique. The cloning then follows the process of SCNT, using a suitable surrogate female, typically a close living relative. Eg a host for cloning a woolly mammoth would be a female elephant, most likely an African elephant, as they are larger than Asian elephants so closer in size to the extinct mammoths. Pyrenean ibex The first attempt at de-extinction was made in 2003 when scientists cloned the Pyrenean ibex which had become extinct in 2000. The clone only lived a few minutes, but recent advances (including the ability of scientists to manufacture DNA to order) means the chances of the clone surviving are greatly improved. In January 2013, Australian scientists announced they had developed embryos of the gastric brooding frog which had become extinct in the 1980’s. Note – just because we can bring species back from the dead, doesn’t mean that we should. There are many potential biological implications concerned with cloning extinct species. Gastric Brooding frog Readings The Microbial World: Biology and Control of Crown Gall (Agrobacterium tumefaciens) Plant Transformations using Agrobacterium tumefacians Videos Ti-Plasmid animation Gene Transfer in Plants using Ti-Plasmid