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Packaging HIV- or FIV-based Lentivector Expression Constructs & Transduction of VSV-G Pseudotyped Viral Particles
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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
Lentiviral expression vectors are the most effective vehicles for stably expressing different effector molecules or reporter constructs in dividing and non-dividing mammalian cells and whole organisms. Here we provide a protocol on how to package lentivector expression constructs in pseudoviral particles and to transduce target cells using the pseudoviral particles.
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
