Stable Cell Line Generation Services - VectorBuilder

Stable cell lines with point mutations are generated using a CRISPR-based approach wherein a target site specific gRNA is introduced into the cells along with ... PleaseWait... Login or Register {[account.info.display]} {[account.info.username]} | Logout  |  DistributorPortal | MyTerritory Users OrderRequests Orders Shipments BundledShipments PriceInquiries DesignRequests FinancialInformation GroupInvoice OrderBasedInvoices MyAccount MySavedDesigns Vectors ServiceProposals MyInquiries PriceInquiries {[messages.price_inquiry.length]} DesignRequests {[messages.design_request.length]} MyBusiness Orders {[messages.project_progress_inquiry.length]} Quotes Invoices OrderRequests StoreCredit MyInformation MessageCenter {[messageTotal]} Profile Address CreditCard YourShoppingCartisempty Youcanfillitinseveralways: Afterdesigningavector,addittoyourcart. Retrieveyoursavedvectorsbygoingtomenuitem "MyAccount"> "MySavedDesigns-Vectors", andaddselectedvectorstoyourcart. 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Subtotal ({[cart.selectedQty]} {[cart.selectedQty===1?('item'|boatFixI18n|translate):('items'|boatFixI18n|translate)]}): ViewCart&Checkout {[site.name]} DesignVector DesignMyVector SendDesignRequest FindPopularVectorsbyGene RetrieveVectorInformation RetrieveServiceProposal Products&Services MolecularBiologyServices OnlineVectorDesign VectorCloning LibraryConstruction BACRecombineering PlasmidDNAPreparation InVitroTranscriptionServices Mutagenesis(forVectors) StableCellLineGeneration ViralVectors&VirusPackaging Lentivirus Adeno-AssociatedVirus(AAV) Adenovirus MMLVRetrovirus MSCVRetrovirus Baculovirus VesicularStomatitisVirus(VSV)  NEW HerpesSimplexVirus(HSV)Vectors  NEW Products shRNALibraries  HOT Dual-gRNACRISPRLibraries  HOT AAVVirus-LikeParticles(VLPs)  New AAVSerotypeTestingPanel   EGFPandHiExpress™LuciferaseIVTmRNA  New VBUltraStableCompetentCells   FeaturedOfferings mRNAGeneDeliverySolutions NEW AAVCapsidEvolution NEW AAVBiodistributionProfiling NEW CRISPRGenomeEditingSolutions InducibleGeneExpressionSolutions shRNAGeneKnockdownSolutions shRNA(3+1)VirusPackaging PurchaseStoreCredit COVID-19CoronavirusSolutions LentivirusPseudotypedwithCoronavirusSpike(S)Protein VSVPseudotypedwithCoronavirusSpike(S)Protein AnimalModelsforCoronavirusResearch GMPManufacturing Overview ProcessDevelopment AnalyticalDevelopment PlasmidManufacturing VirusManufacturing GMPFacilities GMPManufacturing Overview ProcessDevelopment AnalyticalDevelopment PlasmidManufacturing VirusManufacturing GMPFacilities Tools SequenceAlignment SequenceDotPlot shRNATargetDesign CodonOptimization GCContentCalculator DNASecondaryStructure DNAReverseComplement DNATranslation Resources LearningCenter HowtoUseVectorBuilder VideoGallery GuidetoVectorSystems GuidetoVectorComponents GuidetoHostStrains VectorFAQs Documents UserInstructions CertificateofAnalysis MSDS Community TechnicalBulletin Testimonials Citations Support OrderSupport MaterialsSubmissionForm PlaceOrder PayInvoice TrackShipment RetrieveCOA PurchaseStoreCredit Outreach Brochures&Flyers Promotions UpcomingEvents LegalInformation GotoVectorBuilder1.0 AboutUs AboutVectorBuilder OurTeam Newsroom Careers ContactUs Distributors&Marketplaces Home Products&Services StableCellLineGeneration OnlineVectorDesign VectorCloning LibraryConstruction BACRecombineering PlasmidDNAPreparation InVitroTranscriptionServices Mutagenesis(forVectors) StableCellLineGeneration Lentivirus Adeno-AssociatedVirus(AAV) Adenovirus MMLVRetrovirus MSCVRetrovirus Baculovirus VesicularStomatitisVirus(VSV) HerpesSimplexVirus(HSV)Vectors shRNALibraries Dual-gRNACRISPRLibraries AAVVirus-LikeParticles(VLPs) AAVSerotypeTestingPanel EGFPandHiExpress™LuciferaseIVTmRNA VBUltraStableCompetentCells mRNAGeneDeliverySolutions AAVCapsidEvolution AAVBiodistributionProfiling CRISPRGenomeEditingSolutions InducibleGeneExpressionSolutions shRNAGeneKnockdownSolutions shRNA(3+1)VirusPackaging PurchaseStoreCredit LentivirusPseudotypedwithCoronavirusSpike(S)Protein VSVPseudotypedwithCoronavirusSpike(S)Protein AnimalModelsforCoronavirusResearch StableCellLineGeneration VectorBuildercanprovidestablecelllineproductionserviceswithdesiredgeneticmodificationsatunbeatablepricesandrapidturnaround.VectorBuildercandesignandgeneratecustomizedvectorsbasedonyourexperimentalneedsandthecharacteristicsofyourspecificcelllines.Chemicaltransfection,electroporationorviraltransductionareemployedasappropriatetointroducethedesiredDNAchangesintothetargetcells.Followingstandarddrugselection,polyclonalcellpopulationsormonoclonalcelllinesaresub-cultured,verifiedanddeliveredtoyoureadyforuse.Allcellsareguaranteedwithdesiredgenotypes,contaminant-freeandauthenticatedaccordingtoATCCstandards. Typesofstablecelllinesoffered Overexpressionstablecelllines shRNAknockdownstablecelllines Tetinduciblegeneexpressionstablecelllines CRISPRstablecelllines(includesgeneknockout,knockinandpointmutation) ServiceDetails TechnicalInformation HowtoOrder FAQ ServiceDetails Priceandturnaround StableCellLineModel Approach Deliverable Price(USD) Turnaround Geneoverexpression Lentivirus-based Pooledcells From$3,199 9-15weeks Singleclone From$5,199 12-20weeks shRNAgeneknockdown Lentivirus-based Pooledcells From$5,699 8-13weeks Singleclone From$7,699 11-18weeks Tetinduciblegeneexpression Lentivirus-based Pooledcells From$3,599 10-16weeks Singleclones From$5,899 13-21weeks Geneknockout RegularplasmidCRISPR/Cas9-based Singleclone From$5,999 10-16weeks LentiviralCRISPR/Cas9-based Singleclone From$6,999 10-17weeks Geneknockin RegularplasmidCRISPR/Cas9-based Singleclone From$10,099 14-19weeks Pointmutation RegularplasmidCRISPR/Cas9-based Singleclone From$9,199 13-19weeks TechnicalInformation Geneoverexpression VectorBuildercangeneratestablecelllinesoverexpressingyourgene(s)ofinterest(GOIs)tohelpyouovercomevariationsassociatedwithtransienttransfection. Overexpressionstablecelllinesaregeneratedbylentiviraltransductionfollowedbystandarddrugselection.TheGOIexpressionlevelisvalidatedbyRT-qPCR.Additionally,aseriesofstandardQCassayssuchassterilitytestsandmycoplasmadetectionareperformedforreleasingthefinalcelllineproducts.OthervalidationassayssuchasWesternblotandimmunocytochemistrycanbeperformeduponrequest. shRNAgeneknockdown VectorBuildercangenerateshRNAknockdownstablecelllinesforapplicationsrequiringlong-termknockdownofyourGOI(s).ToensureefficientknockdownofyourGOI(s),thetargetcellsareseparatelytransducedwiththreelentivirusescarryingthetop3shRNAsbasedonknockdownscore.ThecelllineshowingthebestknockdownefficiencyvalidatedbyRT-qPCRwillbedelivered.Additionally,aseriesofstandardQCassayssuchassterilitytestsandmycoplasmadetectionareperformedforreleasingthefinalcelllineproducts. Tetinduciblegeneexpression VectorBuildercancustombuildTetinduciblegeneexpressionstablecelllineswithminimalbackgroundexpressionandhighinductionofyourGOI(s).Additionally,wecangenerateTetregulatoryproteinexpressionstablecelllines(e.g.rtTA,tTS/rtTA,etc.)whichcanthenbetransfected/transducedwithplasmids/virusescarryingTREdrivenGOI(s)forflexibleexperimentaldesignandreliableTetinduciblegeneexpression.ForTet-Onstablecelllines,inductionoftheGOIisvalidatedbyRT-qPCR.Additionally,aseriesofstandardQCassayssuchassterilitytestsandmycoplasmadetectionareperformedforreleasingthefinalcelllineproducts. Geneknockout VectorBuildercangeneratestablecelllineswithpermanentknockoutofyourGOI(s).StableknockoutofGOI(s)isachievedusingaCRISPR-basedapproach,whereinapairofgRNAstargetingtheGOI(s)isintroducedintothetargetcellsalongwithCas9.Thisleadstothegenerationoftwocutsonthetargetgene.Attemptsbycellstorepairthebrokenendsofthetwocutsiteswouldtypicallyleadtoalargedeletionspanningthetwosites.Thetwocutsitesaredesignedtoflankafunctionallyimportantregionofthetargetgenesuchthatadeletionspanningthemwouldmostlikelyleadtothelossofgenefunction. Dependingonthecelltype,eitherlentiviralorregularplasmidCRISPRvectorsmaybeusedforachievingtargetgeneknockout.ThefinalcelllineisevaluatedbySangersequencingofthetargetedlocustoconfirmknockout.Additionally,aseriesofstandardQCassayssuchassterilitytestsandmycoplasmadetectionareperformedforreleasingthefinalcelllineproducts. Geneknockin VectorBuildercangeneratestablecellslinewithpermanentknockinofyourGOI(s)atdesiredgenomictargetsites.StableknockinofGOI(s)isachievedbyaCRISPR-basedapproachwhereinatargetsitespecificgRNAisintroducedintothecellsalongwithCas9andadonorvectorwhichservesasthetemplateforHDR-mediatedgeneknockin. ThefinalcelllineisevaluatedbySangersequencingofthetargetedlocustoconfirmknockin.Additionally,aseriesofstandardQCassayssuchassterilitytestsandmycoplasmadetectionareperformedforreleasingthefinalcelllineproducts. Pointmutation VectorBuildercangeneratestablecelllinesharboringdesiredpointmutationsatgenomictargetsitesofinterest.StablecelllineswithpointmutationsaregeneratedusingaCRISPR-basedapproachwhereinatargetsitespecificgRNAisintroducedintothecellsalongwithCas9andasingle-strandeddonoroligonucleotide(ssODN)whichservesasthetemplateforHDR-mediatedinsertionofthepointmutation. ThefinalcelllineisevaluatedbySangersequencingofthetargetedlocustoconfirmthepresenceofthepointmutation.Additionally,aseriesofstandardQCassayssuchassterilitytestsandmycoplasmadetectionareperformedforreleasingthefinalcelllineproducts. HowtoOrder SendDesignRequest FAQ CanVectorBuilderQCfinalcelllinesbyassayssuchasWesternblot,immunocytochemistryandSouthernblot? StablecelllinesgeneratedbyVectorBuilderundergostringentQCprocedureswhichtypicallyincludestandardassayssuchassterilitytestsandmycoplasmadetectionalongwithappropriateassaystovalidatethegenotypeofthefinalcelllinesdependingonthetypeofstablecellline.Forgeneoverexpression,shRNAgeneknockdownandTetinduciblestablecelllinesRT-qPCRisperformedforvalidatingectopicGOIexpression,lossofGOIexpressionandinductionofGOIexpressioninthefinalcellline,respectively.WhereasforCRISPRstablecelllines(geneknockout,knockinandpointmutation),Sangersequencingofthetargetedregionisperformedtovalidatethepresenceofthedesiredgenomiceditinthefinalcellline. InadditiontothestandardQCassaysmentionedabove,VectorBuildercanperformWesternblot,immunocytochemistryandSouthernblotanalysesongeneratedstablecelllinesuponrequest.Theseassaysareavailableasadd-onservicesandcanbeofferedafterevaluatingthefeasibilityofrequestedassaysforthecorrespondingstablecelllines.However,VectorBuilderdoesnotnecessarilyguaranteetheresultsoftheadd-onvalidationassays,becausesomeassaysheavilyrelyontheavailabilityofvalidatedantibodiesandsuitabletestingsystem(e.g.cellline,etc.). HowdoesVectorBuilderdefineturnaroundtimeforstablecelllines? Forstablecelllinegenerationservices,VectorBuilderdefinesestimatedturnaroundasthetimefromproductioninitiationtocompletion.ProductioncompletionisachievedoncethefinalcelllinepassesQCandisreadytobeshippedout.Turnaroundtimedoesnotincludewaitingtimeforanycustomer-suppliedmaterials(e.g.celllinesortemplateDNAforvectorconstruction),QCofsuchmaterials,andtransittimeforshippingfinaldeliverablestothecustomer. 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