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Verpakking HIV-of FIV-based Lentivector expressie constructen en transductie van VSV-G pseudogetypeerde virale deeltjes
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Chapters
- 00:05Title
- 01:00Intro
- 02:28Background on Efficient Lentiviral Packaging
- 03:31Transfecting 293TN Cells with Packaging Plasmids and Expression Constructs
- 05:47Concentrating the Viral Supernatant
- 07:36Transduction of HT1080 Control Cells for Viral Titering
- 09:21Successful Transfection and Transduction
- 10:19Conclusion
Lentivirale expressievectoren zijn het meest effectief voertuigen stabiel tot expressie verschillende effector-moleculen of reporter-constructen in delende en niet-delende zoogdiercellen en hele organismen. Hier bieden we een protocol over hoe lentivector expressie constructen te verpakken in pseudoviral deeltjes en target cellen met behulp van de pseudoviral deeltjes transduceren.
Tags
HIV-based Lentivector Expression ConstructsFIV-based Lentivector Expression ConstructsTransductionVSV-G Pseudotyped Viral ParticlesPackaging Lentiviral Expression ConstructsTransfection EfficiencyTransient ExpressionPseudoviral ParticlesDifficult-to-transfect CellsPrimary CellsStem CellsDifferentiated CellsLentiviral DeliveryCellular ResponsesChemical TransfectionsTransfection Reagent ToxicityTarget CellsGenomic DNA IntegrationStable ExpressionSmall Hairpin RNA (shRNA)CDNAMicroRNAReporter GeneSelectable MarkerPuromycinGFPPseudoviral ReplicationViral Structural GenesLong Terminal Repeats (LTRs)Lentiviral Packaging
