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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.

Publications from our lab describing LeGO-Vectors or RGB-Marking are listed on the main page.

The following figure shows 293T cells 10 days after transduction with LeGO Vectors.

LeGO-Cer

LeGO-G

LeGO-V

LeGO-T

LeGO-C

{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)