CSB349L17 - GFP attached to a 3 UTR inject into cells and...

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GFP attached to a 3’ UTR inject into cells and monitor it using a reporter. Nuclear Reprogramming refers to the global changes in gene expression in a nucleus. The interest in this field is to understand how these changes take place and what is the consequence to the cell.
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Has been studied trhough nuclear transplants and taking two different cell types and fusing them together and find out how the two nucleis interact (heterokaryons). Can study expression of genes from one of the nuclei or the other in the context of the cell. Can occur in pathological conditions. One of the sets of nuclei starts to spit out chromosomes and you start to lose componenets and one nuclei is rendered inactive and you get weird changes in ploidy. It is possible that factors from one nucleus could be secreted and interact w/ factors from another nuc,. .cross talk. Focus has been on chromatin changes, epigenetic etc. which are essential to reprogram gene expression pattern of a nucleus. Human fibroblasts are fused together with one muscle fibre from a mouse (long cells that look like they are multinucleated). After the cells fuse, what’s the gene expression pattern? Normally fibroblasts in humans have a lot of genes that are not expressed. E.g is NCAM which does not normally express. But in a muscle context, the human cells express NCAM. The reason they can tell the difference b/w the two is that they found an antibody which only fuses to the human NCAM. The conclusion is that the muscle environment reprogrammed the human fibroblast to do something it wouldn’t normally do.
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Probing the genome in more sophisticated ways. Used cell fusion, creating fancy transgenic. BJ human normal diploid cells transfected w/ construct that
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This note was uploaded on 12/03/2010 for the course CSB CSB349 taught by Professor V.tropepe,a.moses during the Fall '10 term at University of Toronto- Toronto.

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CSB349L17 - GFP attached to a 3 UTR inject into cells and...

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