BIOC14 : Lec 9 : Article 1 Discussion - Gene targeting in mice

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Biological Sciences
Patrick Mc Gowan

BIOC14Lec 9Article Discussionst Gene targeting in mice functional analysis of the mammalian genome of the 21centurySlide 1Mario Capecchifirst person to figure out how to make genetically manipulated mouseMouse as a model organismTheir genome is similar to human genomeIn the cases where human genes arent found in mouse models ex 5HTT repeat differences seen in humans we can use these genetic manipulation techniques we can introduce those mutations into a mouse to look at the effects Similar kinds of diseases are caused by same kinds of mutations in mice ashumans mice are easy to maintain in labshort breeding cycle can breed everymonthMicecornerstone forbehavioural genetics research w rodents but rats not so much ratsgreat models to study beh in rodents but not as easy to use in transgenic study 4 reasons that arent clearSlide 2Transgenic approaches nowhighly advanced 2 basic onesoDNA microinjection into single cell embryos to produce mice containing multiple copies of randomly inserted DNAoMore selective approaches of using homologous recombination to target specific genomic lociKnockout mousegenes absent knockingenes replacedSlide 3DNA mircoinjection into singlecell embryooFertilized onecell egg obtained from female given doses of estrogen to get her into a superovulating statethis femalemated w stud maleoIsolate fertilized eggsoInject purified DNA into the male pronucleus of fertilized egg using microneedle male pronucleus bc slightly closer to oocyte surface and easier to visualize but can be done w female pronucleus alsooOocytes 2030 transferred into uterus of pseudopregnant femalegoing thru hormonal changes that are associated w pregnancy therefore body is receptive to implantation of a fertilized embryo but this female had been mated w a castrated male therefore this male didnt seminate and just created a pseudopregnant female
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