Lentiviral Vectors: design, production, and titration

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Lentiviral Vector Systems 5’LTR Ψ SD RRE cPPT wPRE SIN3’LTR 8 KB DU3 R U5 3 rd Gen Constitutive Promoter: RSV CMV Packaging Region Rev responsive element Splice Donor Site central polypurine tract Both cPPT and wPRE increase Transduction efficiency and transgene expression ∆U3 R U5 woodchuck hepatitis virus posttranscriptional regulatory element pHR’ SIN-18 Rev is essential for viral replication Binds mRNAs removing them from splicesome = full-length and partially spliced
Lentiviral Vector Systems 5’LTR Ψ SD RRE cPPT hCMV-P eGFP wPRE SIN3’LTR DU3 R U5 Reporter Vector RNA Polymerase II Constitutive: CMV SV40 hEFp PGK Tissue Specific Reporter ∆U3 R U5
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: Packaging plasmids 4 th generation?
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 and 24: Control HEPES - Transfection BES -
Page 25 and 26: Titer Analysis Possibilities: Virus
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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