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Emballagen HIV-eller FIV-baserede Lentivector ekspressionskonstruktioner og transduktion af VSV-G-pseudotypet virale partikler
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Capítulos
- 00:05Título
- 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 ekspressionsvektorer er de mest effektive køretøjer til stabilt at udtrykke forskellige effektormolekyler eller reporterkonstruktioner i delende og ikke-delende mammale celler og hele organismer. Her giver vi en protokol om, hvordan man pakker lentivector ekspressionskonstrukter i pseudoviral partikler og transducere målceller ved hjælp af de pseudoviral partikler.
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
