基因诊断与基因治疗课件

基因诊断与基因治疗Part1:GeneticdiagnosisandDNAfingerprintsa.Huntington’sDisease(HD)b.CysticFibrosis(CF)c.Sickle-CellHemoglobinMutationd.VNTR基因诊断与基因治疗a.TestingfortheexpandedtrinucleotiderepeatregionsinthehuntingtingenethatareresponsibleforHuntington’sdiseasebyPCRb.DetectionofaMutantGeneCausingCysticFibrosisThegenewasisolatedbyLap-CheeTsui(徐立之)in1989基因诊断与基因治疗Allelespecificoligonucleotide(ASO)ProbeHybridizationsDetectionofaMutantGeneCausingCysticFibrosisbySouthernBoltAnalysis基因诊断与基因治疗c.MolecularDiagnosisOfHumanDiseasesDetectionofthesickle-cellhemoglobin

基因诊断与基因治疗d.DNAFingerprintsVariablenumbertandemrepeats(VNTR)基因诊断与基因治疗DNAfingerprintsAndPaternityTests基因诊断与基因治疗DNATestsandtheMysteryoftheDuchessAnastasia基因诊断与基因治疗PedigreeofCzarNicholasII

基因诊断与基因治疗MitochondrialDNA基因诊断与基因治疗PositionVariablenucleotidesinmitochondrialDNA123456AnnaAndersonManahanCCTTCTCarlMaucher(GrandnephewofFranzisca)CCTTCTDukeofEdinburgh(GrandnephewofAlexandra)TTCCTCForensicapplicationOfDNAfingerprints基因诊断与基因治疗Part2:GeneTherapy

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

Avectordeliversthetherapeuticgeneintoapatient’stargetcellThetargetcellsbecomeinfectedwiththeviralvectorThevector’sgeneticmaterialisinsertedintothetargetcellFunctionalproteinsarecreatedfromthetherapeuticgenecausingthecelltoreturntoanormalstate

GeneTherapyisExperimental

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

基因诊断与基因治疗MajorityareTrails

基因诊断与基因治疗VivotoVitro

基因诊断与基因治疗

Researchersmayuseoneofseveralapproachesforcorrectingfaultygenes:Anormalgenemaybeinsertedintoanonspecificlocationwithinthegenometoreplaceanonfunctionalgene.Thisapproachismostcommon.（基因增补）2.Anabnormalgenecouldbeswappedforanormalgenethroughhomologousrecombination.(基因置换)3.Theabnormalgenecouldberepairedthroughselectivereversemutation,whichreturnsthegenetoitsnormalfunction.（基因矫正）4.Theregulation(thedegreetowhichageneisturnedonoroff)ofaparticulargenecouldbealtered.（基因失活）2.GeneTherapy:ApproachesWhatGenetherapycanAchieve

基因诊断与基因治疗GoalofGenetherapy

基因诊断与基因治疗DeliveringdesiredGenes

基因诊断与基因治疗GeneTherapyCorrects

3.StepsinGeneTherapy

基因诊断与基因治疗AAV-Adeno-associatedvirus（1）ViralGeneTherapy:anoverview基因诊断与基因治疗ManipulationcorrectstheDefectiveGenes

基因诊断与基因治疗GeneTherapydeliversProteins

基因诊断与基因治疗a.AdenovirusVector

基因诊断与基因治疗AnADAgene-transfervectorAdenosinedeaminase-deficientseverecombinedimmuno-deficiencydisease

基因诊断与基因治疗Adeno-associatedviruses

基因诊断与基因治疗AdenovirusVector基因诊断与基因治疗HumanGeneTherapyUsingAnAdenovirusVector基因诊断与基因治疗AdenovirusVectorInjection基因诊断与基因治疗LimitationofDirectGeneInduction

b.RetrovirusVector基因诊断与基因治疗Retroviruses基因诊断与基因治疗RetrovirusandGeneTherapy基因诊断与基因治疗例如：小鼠白血病病毒（Mo-MLV）基因组结构：

LTRLTRψgagpolenvU3RU5U3RU5

U3=增强子R=启动子U5=转录起始位点。Ψ=病毒包装信号gag=核心抗原pol=逆转录酶env=外壳蛋白。

Safety

基因诊断与基因治疗MakingsafeProtocols

基因诊断与基因治疗Celltypespecificity

基因诊断与基因治疗Lentivirus基因诊断与基因治疗VectorsdelivertheGeneticMaterials

基因诊断与基因治疗c.Nonviralapproach

基因诊断与基因治疗NonviralVectors:

Liposome'slessImmunogenic

基因诊断与基因治疗Nanoengineeredsubstances

基因诊断与基因治疗TransfectionandNanoengineering

基因诊断与基因治疗CreatingNewChromosome

基因诊断与基因治疗（2）Antisensetherapy

基因诊断与基因治疗Antisensetherapy

基因诊断与基因治疗AntisenseRNAandGeneTherapy

DNADNA

RNARNA

反义RNA

阻断表达

基因诊断与基因治疗AntisenseRNA(oligodeoxyribonucleotide,ODNortriplex-formingoligo,TFO)andGeneTherapyTargetedinhibitionofgeneexpression基因诊断与基因治疗（3）RNAiandGeneTherapy基因诊断与基因治疗（4）RibozymeandGeneTherapy“hammerhead”ribozymesRecognitionsequences基因诊断与基因治疗SelfSplicingIntronRibozymes基因诊断与基因治疗4.DevelopmentsinGeneTherapy

FirstApprovedGeneTherapy

基因诊断与基因治疗Whatdidtheydo

基因诊断与基因治疗Asuccessstory

基因诊断与基因治疗ADAdeficiencywasselectedforthefirstapprovedhumangenetherapytrialforseveralreasons

基因诊断与基因治疗GeneTherapy:

UsesAIDSVirustoFightAIDS

基因诊断与基因治疗Thecurrentstatusofgenetherapyresearch

Littleprogresshasbeenmadesincethefirstgenetherapyclinicaltrialbeganin1990.In1999,genetherapysufferedamajorsetbackwiththedeathof18-year-oldJesseGelsinger.基因诊断与基因治疗JesseGelsinger’sDeath

Jessewasparticipatinginagenetherapytrialforornithinetranscarboxylase(鸟氨酸转羧酶)deficiency(OTCD).Hediedfrommultipleorganfailures4daysafterstartingthetreatment.Hisdeathisbelievedtohavebeentriggeredbyasevereimmuneresponsetotheadenoviruscarrier.基因诊断与基因治疗FDA’sAction

AnothermajorblowcameinJanuary2003,whentheFDA(USA)placedatemporaryhaltonallgenetherapytrialsusingretroviralvectorsinbloodstemcells.基因诊断与基因治疗FDA’sAction

FDA(USA)tookthisactionafteritlearnedthatasecondchildtreatedinaFrenchgenetherapytrialhaddevelopedaleukemia-likecondition.基因诊断与基因治疗TwoChildren’sCondition

BoththischildandanotherwhohaddevelopedasimilarconditioninAugust2002hadbeensuccessfullytreatedbygenetherapyforX-linkedseverecombinedimmunodeficiencydisease(X-SCID),alsoknownas"bubblebabysyndrome."基因诊断与基因治疗BRMAC’sAttitude

FDA'sBiologicalResponseModifiersAdvisoryCommittee(BRMAC)metattheendofFebruary2003todiscusspossiblemeasuresthatcouldallowanumberofretrov