Lentivirus - 4. page

In HIV, pseudotyping is used to change the tropism of the virion.  In other words, pseudotyping changes the way the Lentivector targets a specific cell for infection.  In wild type […]

Though there are several virus delivery systems available today for gene therapy, lentivirus is considered one of the best methods.  Several features make lentivirus preferable especially the ability to infect […]

Family Genus Species Vector References Rhabdoviridae Vesiculovirus Vesicular stomatitis virus (Indiana virus) HIV-1 [Akkina, 1996][Naldini, 1996][Reiser, 1996] HIV-2 [Poeschla, 1998a] FIV [Poeschla, 1998b] EIAV [Olsen, 1998] BIV [Berkowitz, 2001a] JDV […]

Feature 2nd Generation 3rd Generation Transfer Plasmid Can be packaged ONLY by a second generation packaging system that includes TAT Can be packaged by both 2nd and 3rd generation packaging […]

Vector and region amplified Primers and probes dedicated for the amplification of lentiviral or transgenic DNA sequences Primers and probes dedicated for the amplification of internal control DNAs (cellular genome […]

1. Extract RNA using a commercial kit 2. DNase treatment using RNase free DNase 1U/ul from Promega to digest residual plasmid DNA 3. Use ABI TaqMan One-Step RT-PCR master mix […]

1. Make two primer probe sets U5-Psi: U5-Psi forward: TGTGTGCCCGTCTGTTGTGT U5-Psi reverse: GAGTCCTGCGTCGAGAGAGC U5-Psi probe: FAM-CAGTGGCGCCCGAACAGGGA-TAMARA WPRE: WPRE forward: CCGTTGTCAGGCAACGTG WPRE reverse: AGCTGACAGGTGGTGGCAAT WPRE probe: FAMTGCTGACGCAACCCCCACTGGT-TAMARA 2. Purify RNA from […]

Ultracentrifuge: Beckman LE 80K Rotor: SW28 Tubes: #326823 autoclaved (polyallomer) 1. Pool all viral media into and filter through a 0.45μm HV Durapore membrane (Millipore #SCHVU01RE). 2. Place tubes into […]

Before Starting: Required equipments and consumbles: 1. SW-28 rotor with buckets and caps (Kept in 4C Refrigerator in Centrifuge Room) 2. SW-28 thin walled ultra clear centrifuge tubes (Beckman # […]