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JOUR 3440 (11)
Lecture

Transgenic Organisms - Section 4

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Department
Journalism
Course
JOUR 3440
Professor
billpohajdak
Semester
Winter

Description
08092014000200Lecture213122014HumaninsulinKnowhowtodoaknockoutPuttingintopluripotentandtotipotentKnockoutYouremovethegeneyouwanttoknockoutYoumodifyitgeneticallyoutsideoftheorganismYouputinamarkerinoneoftheexonsorpartofthecodingregionNeomycinisthemarkerItisaversionthatwillkillhumancellsifyouaddG418ItsaselectionIfyoudisruptthegenethenmRNAisdisruptedandproteinproductiongetsmessedupUsuallydesignedtoknockoutOutsideofthegeneyoutagonanotherpieceofsyntheticDNAHerpessimplexvirusThymaninekinaseisdifferentandtotreatherpestheymadeadrugthatinhibitsthisThedrugisganciclovirItlookslikeguaninebutthesugarpartisonlyhalfSowhathappensistheherpesthymaninekinaseItrecombineswiththehostDNAbutitsnotintherightplaceCrossoverbetweenregionsofDNAthatareidenticalhomologousrecombinationThisiswhywehavedifferentphenotypicmarkersBecausethedisruptionisthereournewgeneifthecrossoveroccurredwithhomologousrecombthenewgenewillbeknockedoutItwonthappenonbothchromosomesThyminekinasegetslostafterhomologousrecombinationThesecellsareresistanttoneomycinDuringnonhomologousrecombinationthecellsareresistanttoneomycinbuttheyaresensitivetoganciclovirWecankillthiscellbyaddingganciclovirTransgenicMiceMicehavemoregenesthanwedobut95areidenticalinfunctionStudiesusingtransgenicmiceRescueofmutantsMiceasbioreactorssynthesisofhumanmonoclonalantibodiesStructurefunctionrelationshipsMousemodelsofhumandiseaseAnalysisofcellularandmolecularmechanismsunderlyingpathogenesisandfortestingdrugregimesandorgenetherapiesProductionoftransgenicmiceTakesabout5monthstomaketransgenictomakeknockoutittakes23yearsPronuclearDNAmicroinjectionCloningembryosisreallyeasybyinjectingtheeggimplantingfetusesorblastocystsisalotharderLecture223142014TransgenicMiceThegrowthhormonegenehasbeenengineeredtobeexpressedathighlevelsinanimals1983andtheresultwasbiganimalsColorfulMiceTheywanttomaketissuethatisfluorescentorpartsTheGFPgotthemostnewscamefromjellyfishAddgenetopromoteranditdoesnthurtorganismFurisnotgreenbecausefurisnotmadeofcellsTheydecidedtomakeredmiceComesfromcoralcanuseittowatchwhatacelldoesinanorganismTookmultiplecopiesinthesamegeneconstructandfloxeditbluegreenandred3copiesandthenrandomlycutout123or4copiesAndthentheywouldgetarangeTheylookedatindividualneuronsandcallthemBrainbowmicebecausetheycandothisOnlyproblemistheydontknowwhatelsetodowiththesemiceHuMAbmouseisusedinwomenwithbreastcancerIfyoumaketheantibodiesinamousethenyoucanmakehumanantibodiesPartialhumanmicealmostallmicethatareusedareidenticaltoeachotherifyoutakeamouseandcrossitbrothersistermateanddoitfor20generationstheyareallidenticalTheywillacceptanykindoftransplantetcyoucangeneticallymakeyourgeneonabackgroundofamousethatisallthesameHepatitisCmouse
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