What Is Gene Floxing and Why Is It Used by Scientists Today?

Floxing, or flanking by loxP, is a process used by geneticists by which a specifically targeted gene is flanked by two loxP sites. Floxing February2,2021|ingenious|Cre-lox WhatIsGeneFloxingandWhyIsItUsedsoFrequentlybyScientistsToday? Floxing,orflankingbyloxP,isaprocessusedbygeneticistsbywhichaspecificallytargetedgeneisflankedbytwoloxPsites.Althoughseeminglysimple,thisprocessisrelativelynewandquitecomplex.Inmostcases,genetargetingwiththeuseoffloxingisenhancedandcatalyzedthroughaprocessknownasCre/loxrecombination.Thiscanleadtotheproductionofappropriategeneticmodelsforthestudyofvariousgeneticdiseases,aswellasdiseasesthatcanbetriggeredbythemutationorinactivationofacertaingene.Manystudieshaveshowntheeffectivenessofusingfloxedanimalmodels–especiallymousemodels–inordertoobtainconditional,tissue-specificmodificationsaswellasthealterationofvariousgeneexpressions. Inessence,throughtheuseoffloxingandCrerecombinase,itispossibletospliceageneentirelyoutofthegenome,inverttheorientationofthegenebyremovingitandsplicingitbackinwiththeoppositeorientation,andreplacethegenewithafloxedDNAsequencethatoriginatedelsewhere.AlloftheseoptionsareavailableonceCrerecombinaseisexpressedandbecomesactivewithinacellwithafloxedgene. TheMainAdvantagesofGeneFloxing Oneofthegreatestadvantagesoffloxingisthatitallowsfordetailedinvivostudyofabnormalitiesanddiseasesassociatedwithgeneticmodifications.Duringthedevelopmentoffloxedmousemodels,geneknockoutsandotheralterationscanbetriggeredbycertainprocessesatspecifictimes.ThisactionismadepossiblebytheuseoffloxingtogetherwiththeCre/loxrecombinationsystem,allowingforthegenerationofbetter,morepreciseanimalmodels.Thedevelopmentofdrugsandgenetictreatmentscanbehighlydependentonnewbreakthroughsandresultsthatarefoundduringtheinvivostudyoffloxedmousemodels.Bettermodelscanacceleratethisprocess,astheyallowresearcherstoobtainmoreaccurateandpracticalresults. Tissue-specificknockoutsusingCre/loxrecombinationandfloxedmicecanpreventmanyoftheproblemsthatoccurduringconventionalknockoutmutations.Thesecanincludeseriousconcernssuchasincreasedembryoniclethalityandunwantedinconsistenciesinthegeneknockoutprocess.Asaresult,whenevergreateraccuracyoratissue-specificrecombinationisneeded,mostexpertswillrecommendtheuseoffloxedmousemodelsthathavethegeneinquestioninactivatedonlyinaspecifictypeoftissue,suchasthekidneysorliver. Otherusesforthesite-specificrecombinasesystemusinggenefloxingcanalsoincludeconditionalgenetrapping,thecreationofamultipurposeconditionalalleleandtheabilitytofacilitatemoreaccuratemethodsforgenesilencing.Alltheseusescomewiththeirownadvantages,includingthereductionofembryolethalityinthecaseofgenetrappingstudiesandthepossibilitytoinducegenesilencinginlesstime,comparedtoconventionalmethods. Insummary,genefloxingispowerfulintoday’sresearchlaboratoriesasitallowspreciseanddiscreteanalysisofgenefunction.Itsuseinanimalbiologywillbetterallowustodelineatetheexactroleofmultiplecelltypesandatvaryingtimesthroughoutthelifespanofananimal.Forexample,usinggenefloxingwiththealpha-MyHCpromotercausesthefloxedgenetobeinactivatedintheheartalone.Further,theseknockoutscanbeinducible.Inseveralmousestudies,tamoxifenisusedtoinducetheCrerecombinase.Inthiscase,Crerecombinaseisfusedtoaportionofthemouseestrogenreceptorwhichcontainsamutationwithinitsligandbindingdomain.Themutationrendersthereceptorinactive,whichleadstoincorrectlocalizationthroughitsinteractionswithchaperoneproteinssuchasheatshockprotein.TamoxifenbindstoCre-ERanddisruptsitsinteractionswiththechaperones,whichallowstheCre-ERfusionproteintoenterthenucleusandperformrecombinationonthefloxedgene.Thus,thisgeneticresourceisavailabletocontinuetheremarkableprogressmaderecentlyinunderstandinghowcertaingenesfunctiontoregulatedevelopmentandsignalingactivities. Lastupdated:February2nd,2021 Sharethis: Facebook Twitter LinkedIn 100%satisfactionwithingenious’mousemodelservices“Weare100%satisfiedwithiTL’sserviceingeneratingthismousemodelforus.Themodelwascompletedquickly,withtheutmostefficiencyandeachmemberoftheirteamwas[…]JosephA.Fraietta,PhDPerelmanSchoolofMedicine,UniversityofPennsylvaniaReadmoreSimilarArticles