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Homepage of the Lentiviral Gene Ontology Vectors |
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{Lentiviral Gene Ontology Vectors}
The Lentiviral Gene Ontology Vectors are a "multi-color" panel of lentiviral vectors designed according
to the "building blocks" principle. Each vector allows expression of one transgene of interest and/or a
shRNA of choice and, in addition, a fluorescent marker protein for identification and/or selection of
transduced cells. To permit simultaneous analysis of multiple genes, a large spectrum of fluorescent
marker genes has been used, including eGFP/BSD and dTomato/BSD fusion genes, which combine the advantages
of both fluorescent and drug-selectable marker genes. These vectors are useful for concurrent over-expression
as well as suppression of various genes, thus allowing gene function studies in a variety of target cells.
In accordance with their design and envisioned application for analysis of gene networks, we have named our
constructs Lentiviral “Gene Ontology” (LeGO) vectors.
{Principle of Complementation} Third-generation lentiviral vectors do not encode any viral proteins but contain cis-active elements for packaging, reverse transcription and integration. This design not only incorporates safety features, but also provides space for up to 9 kb of foreign sequences. The viral genes necessary for the production of the viral particles (gag/pol, rev and env) have to be supplied in trans, which is done by a co-transfection of the producer cells with 4 plasmids. See also
Naldini et al. (1996),
Dull et al. (1998).
{Reverse Transcription and self-inactivation} This is a scheme of how self-inactivating (SIN) vectors work. On the vector plasmid, enhancer/promoter sequences of the U3 region of the 3'-LTR have been deleted (ΔU3). During the reverse transcription, this "defective" U3-region will be copied to the 5'-end, resulting in a provirus with this enhancer/promoter-deletion in both LTRs. See also
Miyoshi et al. (1998)
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