基因診斷與基因治療課件

基因診斷與基因治療Part1:GeneticdiagnosisandDNAfingerprintsa.Huntington’sDisease(HD)b.CysticFibrosis(CF)c.Sickle-CellHemoglobinMutationd.VNTR基因診斷與基因治療a.TestingfortheexpandedtrinucleotiderepeatregionsinthehuntingtingenethatareresponsibleforHuntington’sdiseasebyPCRb.DetectionofaMutantGeneCausingCysticFibrosisThegenewasisolatedbyLap-CheeTsui(徐立之)in1989基因診斷與基因治療Allelespecificoligonucleotide(ASO)ProbeHybridizationsDetectionofaMutantGeneCausingCysticFibrosisbySouthernBoltAnalysis基因診斷與基因治療c.MolecularDiagnosisOfHumanDiseasesDetectionofthesickle-cellhemoglobin

基因診斷與基因治療d.DNAFingerprintsVariablenumbertandemrepeats(VNTR)基因診斷與基因治療DNAfingerprintsAndPaternityTests基因診斷與基因治療DNATestsandtheMysteryoftheDuchessAnastasia基因診斷與基因治療PedigreeofCzarNicholasII

基因診斷與基因治療MitochondrialDNA基因診斷與基因治療PositionVariablenucleotidesinmitochondrialDNA123456AnnaAndersonManahanCCTTCTCarlMaucher(GrandnephewofFranzisca)CCTTCTDukeofEdinburgh(GrandnephewofAlexandra)TTCCTCForensicapplicationOfDNAfingerprints基因診斷與基因治療Part2:GeneTherapy

1.GeneralConceptsofGeneTherapyGenetherapyistheinsertionofgenesintoanindividual'scellsandtissuestotreatadisease,suchasahereditarydiseaseinwhichadeleteriousmutantalleleisreplacedwithafunctionalone.Althoughthetechnologyisstillinitsinfancy,ithasbeenusedwithsomesuccess.HowItWorks

Avectordeliversthetherapeuticgeneintoapatient’stargetcellThetargetcellsbecomeinfectedwiththeviralvectorThevector’sgeneticmaterialisinsertedintothetargetcellFunctionalproteinsarecreatedfromthetherapeuticgenecausingthecelltoreturntoanormalstate

GeneTherapyisExperimental

Advancesinunderstandingandmanipulatinggeneshavesetthestageforscientiststoalteraperson'sgeneticmaterialtofightorpreventdisease.Genetherapyisanexperimentaltreatmentthatinvolvesintroducinggeneticmaterial(DNAorRNA)intoaperson'scellstofightdisease.

基因診斷與基因治療MajorityareTrails

基因診斷與基因治療VivotoVitro

基因診斷與基因治療

Researchersmayuseoneofseveralapproachesforcorrectingfaultygenes:Anormalgenemaybeinsertedintoanonspecificlocationwithinthegenometoreplaceanonfunctionalgene.Thisapproachismostcommon.（基因增補）2.Anabnormalgenecouldbeswappedforanormalgenethroughhomologousrecombination.(基因置換)3.Theabnormalgenecouldberepairedthroughselectivereversemutation,whichreturnsthegenetoitsnormalfunction.（基因矯正）4.Theregulation(thedegreetowhichageneisturnedonoroff)ofaparticulargenecouldbealtered.（基因失活）2.GeneTherapy:ApproachesWhatGenetherapycanAchieve

基因診斷與基因治療GoalofGenetherapy

基因診斷與基因治療DeliveringdesiredGenes

基因診斷與基因治療GeneTherapyCorrects

3.StepsinGeneTherapy

基因診斷與基因治療AAV-Adeno-associatedvirus（1）ViralGeneTherapy:anoverview基因診斷與基因治療ManipulationcorrectstheDefectiveGenes

基因診斷與基因治療GeneTherapydeliversProteins

基因診斷與基因治療a.AdenovirusVector

基因診斷與基因治療AnADAgene-transfervectorAdenosinedeaminase-deficientseverecombinedimmuno-deficiencydisease

基因診斷與基因治療Adeno-associatedviruses

基因診斷與基因治療AdenovirusVector基因診斷與基因治療HumanGeneTherapyUsingAnAdenovirusVector基因診斷與基因治療AdenovirusVectorInjection基因診斷與基因治療LimitationofDirectGeneInduction

b.RetrovirusVector基因診斷與基因治療Retroviruses基因診斷與基因治療RetrovirusandGeneTherapy基因診斷與基因治療例如：小鼠白血病病毒（Mo-MLV）基因組結構：

LTRLTRψgagpolenvU3RU5U3RU5

U3=增強子R=啟動子U5=轉錄起始位點。Ψ=病毒包裝信號gag=核心抗原pol=逆轉錄酶env=外殼蛋白。

Safety

基因診斷與基因治療MakingsafeProtocols

基因診斷與基因治療Celltypespecificity

基因診斷與基因治療Lentivirus基因診斷與基因治療VectorsdelivertheGeneticMaterials

基因診斷與基因治療c.Nonviralapproach

基因診斷與基因治療NonviralVectors:

Liposome'slessImmunogenic

基因診斷與基因治療Nanoengineeredsubstances

基因診斷與基因治療TransfectionandNanoengineering

基因診斷與基因治療CreatingNewChromosome

基因診斷與基因治療（2）Antisensetherapy

基因診斷與基因治療Antisensetherapy

基因診斷與基因治療AntisenseRNAandGeneTherapy

DNADNA

RNARNA

反義RNA

阻斷表達

基因診斷與基因治療AntisenseRNA(oligodeoxyribonucleotide,ODNortriplex-formingoligo,TFO)andGeneTherapyTargetedinhibitionofgeneexpression基因診斷與基因治療（3）RNAiandGeneTherapy基因診斷與基因治療（4）RibozymeandGeneTherapy“hammerhead”ribozymesRecognitionsequences基因診斷與基因治療SelfSplicingIntronRibozymes基因診斷與基因治療4.DevelopmentsinGeneTherapy

FirstApprovedGeneTherapy

基因診斷與基因治療Whatdidtheydo

基因診斷與基因治療Asuccessstory

基因診斷與基因治療ADAdeficiencywasselectedforthefirstapprovedhumangenetherapytrialforseveralreasons

基因診斷與基因治療GeneTherapy:

UsesAIDSVirustoFightAIDS

基因診斷與基因治療Thecurrentstatusofgenetherapyresearch

Littleprogresshasbeenmadesincethefirstgenetherapyclinicaltrialbeganin1990.In1999,genetherapysufferedamajorsetbackwiththedeathof18-year-oldJesseGelsinger.基因診斷與基因治療JesseGelsinger’sDeath

Jessewasparticipatinginagenetherapytrialforornithinetranscarboxylase(鳥氨酸轉羧酶)deficiency(OTCD).Hediedfrommultipleorganfailures4daysafterstartingthetreatment.Hisdeathisbelievedtohavebeentriggeredbyasevereimmuneresponsetotheadenoviruscarrier.基因診斷與基因治療FDA’sAction

AnothermajorblowcameinJanuary2003,whentheFDA(USA)placedatemporaryhaltonallgenetherapytrialsusingretroviralvectorsinbloodstemcells.基因診斷與基因治療FDA’sAction

FDA(USA)tookthisactionafteritlearnedthatasecondchildtreatedinaFrenchgenetherapytrialhaddevelopedaleukemia-likecondition.基因診斷與基因治療TwoChildren’sCondition

BoththischildandanotherwhohaddevelopedasimilarconditioninAugust2002hadbeensuccessfullytreatedbygenetherapyforX-linkedseverecombinedimmunodeficiencydisease(X-SCID),alsoknownas"bubblebabysyndrome."基因診斷與基因治療BRMAC’sAttitude

FDA'sBiologicalResponseModifiersAdvisoryCommittee(BRMAC)metattheendofFebruary2003todiscusspossiblemeasuresthatcouldallowanumberofretrov