Many products of major therapeutic value can be obtained only from animal cells grown in culture. The aim of this project is to improve the processes currently employed in this sector of biotechnology with special emphasis on the vectors and promoters used and on the quality of post translational modification.
This work concerns the optimalization of linear, parvovirus-based vectors for the transfer and expression of foreign genes in mammalian cells. Existing molecular clones of parvovirus MVM will be modified by site directed mutagenesis of the P4 promoter and/or downstream regulatory sequences, in order to boost expression from this promoter.
A packaging cell line, constitutively expressing capsid proteins, will be constructed for the obtaining stocks of recombinant parvoviruses.
In order to extend the range of systems allowing the sustained propagation of replication-competent MVM-based vectors, new parvoviral and cellular variants will be isolated that produce non-cytotoxic NS proteins or are resistant to the wild-type protein, respectively. The hyperactivity of parvoviral promoters in a number of transformed cells will be exploited to achieve targeted killing of these cells by the products of appropriate genes places under parvoviral control. These can either be toxins (HSV-1 thymidine kinase in combination with acyclovir) or products that stimulate the immune system (cytokines). The interference of parvovirus driven interleukin genes with the formation of tumours in experimental animals will be assessed.
Funding SchemeCSC - Cost-sharing contracts