Lentiviral Vectors: design, production, and titration

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FACs Titer: pLV FACs detects Infected cells, Expressed as Percentage of total pMDLpg.RRE pRSV-Rev pMD.G Infection Transfection Conditioned Medium Transfected 293T cells Infected 293T cells
Titer Analysis Possibilities: Virus Production FACS for gene expression product: Dependent on Promoter activity Constitutive promoter = useful Titers that predict infection rate Tissue specific promoters might not give useful titers Real-Time PCR for integrated viral DNA in host genome Dependent on infection <strong>and</strong> integration into the host genome Real-Time PCR Titers predict infection rate Reverse transcription Real-Time PCR for viral RNA Dependent only on the presence of the viral RNA Does not predict infection rate of the viral particles
Page 1 and 2: Lentiviral Vectors: design, product
Page 3 and 4: Lentiviral Vector System 2. Modific
Page 5 and 6: HIV provirus LTR ψ SD Gag Pro 2 nd
Page 7 and 8: Packaging plasmids 4 th generation?
Page 9 and 10: Lentiviral Vector Systems 5’LTR
Page 11 and 12: Lentiviral Vector System (3 rd gene
Page 13 and 14: Lentiviral Vector System (3 rd gene
Page 15 and 16: 1 2 3 4 5 DU3 R U5 DU3 R U5 DU3 R U
Page 17 and 18: Lentiviral Vector System: Gene Supp
Page 19 and 20: Lentiviral Vector System: Gene Supp
Page 21 and 22: Lentiviral Vector System Transgene
Page 23: Control HEPES - Transfection BES -
Page 28 and 29: % fluorescence 70 60 50 40 30 20 10
Page 30 and 31: Replication Competent Lentivirus (R

Lentiviral Vectors: design, production, and titration

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