Lentiviral Vectors

All vectors are based on the lentiviral shuttlevector FUGW from Lois et al. 2002. In this basic vector we exchanged the promoter and included a multiple cloning site (MSC). For now we are using CMV, CamKII, hSynapsinI, GFAP or hUbiquitin C Promoters for driving the expression of the transgenes.

In a second type of LV vectors we employed different reporter genes together with a self-cleaving 2A peptide from porcine teschovirus-1 (P2A) followed by the MCS. This allows us the bicistronic expression of reportergene and transgene making it possible to carefully characterize virus efficiency.

The third set of viral vectors utilizes the combination of two different Lox sites. The introduction of a reporter gene plus a reverse oriented reporter gene in conjunction with P2A and MSC make these vectors suitable for cell-type specific expression of the transgene in Cre transgenic animals.

For reducing endogenous mRNAs levels (knockdown) we use a viral vector which contains the human RNA polymerase III promoter U6 for driving the transciption of specific shRNAs. A second promoter (hUbiquitin C or hSynapsin1) drives the expression of different reporter genes.

All viruses are generated with the help of two "Helper" plasmids providing packaging and envelope proteins in trans (packaging plasmid pCMV-dR8.9 and envelope plasmid pCMV-VCV-G).

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