Lentiviral Vectors: design, production, and titration

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Replication Competent Lentivirus (RCL) 3 rd Generation <strong>Lentiviral</strong> Vector HIV provirus Wild-Type Virus LTR SD ψ Gag Pro Pol + Replication competent LTR Vif Gag,pol, rev, env, tat R U Source: Carry over from packaging or Envelope plasmids Or Endogenous viruses Tat Rev RRE Env Nef LTR
Replication Competent Lentivirus (RCL) Protocol: 1. Infect 1 million SupT-1 cells with 5 million viral TUs 2. Pass the cells 3 times over 2-3 weeks 3. Test the medium for p24 protein with ELISA kit (commercial) Test Preps + Std
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 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

Lentiviral Vectors: design, production, and titration

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