Talk:CH391L/S12/CounterSelection: Difference between revisions
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Revision as of 18:44, 26 February 2012
- Ben Slater 02:09, 25 February 2012 (EST): Does allelic exchange work in eukaryotes? It seems like it would be awesome for applications like gene therapy, if it allows chromosomal editing at any given site. Another question: how would one synthesize the homologous insert? Thanks!
- Jeffrey E. Barrick 19:08, 26 February 2012 (EST):Here's an attempt at an answer. I'm no expert in this area. You can "transiently transfect" eukaryotic (mammalian) cells in culture by using viruses as vectors. This results in your gene being made, but isn't very stable -- it's like putting an unstable plasmid into a bacterium. You can also generate animals, like mice that have a gene knocked out. I'm not familiar with the exact methods, but it undoubtedly involves using selectable markers. In multicellular eukaryotes, a lot of the difficulty is also in getting the mutation in the germ line, or at least into cells that continue to divide during an organism's lifetime.
- Yi Kou 20:43, 26 February 2012 (EST): I agree with Dr. Barrick's opinion. Clinically, there is not such application so far. Partially because of our comprehension of temporal and spatial arrangement of gene (& its regulation, expression) in multicellular organism is far from satisfying. Regarding temporal aspect, interruption of a gene can easily result in no observation of the expectation, simply because that the gene may be needed on one developmental stage and interruption of it leads to a termination of an observable maturity. For spatial factor, if a pivotal and universally needed gene has got somewhat modification, its phenotypic results can be seen anywhere in the organism, not leading to a reducible and conclusive manner. However, despite these, there have been developed several techniques, either improve "transfect" stability or make the independent variable more "independent" to be studied [1].
- Jeffrey E. Barrick 19:08, 26 February 2012 (EST):Here's an attempt at an answer. I'm no expert in this area. You can "transiently transfect" eukaryotic (mammalian) cells in culture by using viruses as vectors. This results in your gene being made, but isn't very stable -- it's like putting an unstable plasmid into a bacterium. You can also generate animals, like mice that have a gene knocked out. I'm not familiar with the exact methods, but it undoubtedly involves using selectable markers. In multicellular eukaryotes, a lot of the difficulty is also in getting the mutation in the germ line, or at least into cells that continue to divide during an organism's lifetime.
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