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Obtention of receptor and/or transcriptionally targeted adenoviral vectors efficiently and specifically expressing a marker gene in hepatoma, adenocar

A set of adenoviral vectors were designed and engineered such as to provide efficient and specific expression of a marker gene in hepatoma, adenocarcinoma and glioblastoma cell lines, and neuronal and glial primary cell cultures.

For marking studies, the GFP expressing vector AdFK7 was constructed. This vector expresses EGFP under the control of the hCMV promoter.

To allow the easy switch between liver-specific, glia- or neuron-specific, or constitutively regulatable systems, various shuttle plasmids for HC-Ad vector construction were engineered. In a shuttle plasmid, the specific promoter can be exchanged in a single cloning step, and, in addition, the transgene can be exchanged in a single cloning step. The complete cassette then can be removed from the plasmid backbone as a single fragment and is introduced into an HC-Ad shuttle plasmid in a further cloning step. This plasmid then is ready for rescue by transfection of N52.Cre cells together with loxP helper virus.

To allow production of HC-Ad vectors with targeted capsids, different adenoviral helper viruses were engineered. Adenoviral helper viruses with capsids were modified such as to contain targeting ligands in the HI-loop of the knob domain of the adenoviral fiber protein. These targeting ligands include a cyclic RGD peptide, a polylysine peptide and a VCAM1 binding peptide. They allow improved transduction of several primary cell types and of several cancer cell lines. Vectors generated by usage of these helper viruses during production will be used to transduce tumours in vivo or endothelial cells in tumours expressing increased VCAM1.

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University of Ulm
Helmholtz Str. 8/1
89081 Ulm
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