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CORDIS - Forschungsergebnisse der EU
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Safety of genetically engineered retroviruses used for gene transfer

Ziel

The objective of the project is to accumulate basic information regarding cellular and viral mechanisms involved in modulating the behaviour of retroviral vector systems and to use this knowledge as a basis for an assessment of the risk of using retroviral vectors for the production of transgenic animals or for human gene therapy, and to construct safe, efficient and targeted retroviral vector systems.
Developments in the field of retroviral vector mediated gene therapy in humans have demonstrated the importance and requirements for research efforts to determine the safety of genetically engineered retroviruses. Results of work to date follow.

Work is progressing with respect to transfer, expression and genetic stability of murine leukemia virus derived vectors in cultured murine cells. Oligodeoxynucleotide primers for polymerase chain reaction (PCR) amplification and sequence analysis have been designed and synthesized, and nucleotide sequence information of integrated vector proviruses has been obtained. Cell clones with integrated proviruses selected for higher expression have been characterized by Southern hybridization and PCR sequence analysis. Experiments directed towards the analysis of long term stability of expression of integrated proviruses have been initiated and differences among individual proviruses have been observed. Studies analysing the relative influence of promoter-enhancer elements of the vector and of the host chromosomal integration site are continuing. As part of this, studies of the inducibility of individual integrated vector proviruses by the steroid hormone dexamethasone have been completed.

Initial experiments aimed at determining the elements involved in the tissue specific expression of genes in the mammary gland have been completed. Based upon initial findings strategies have been set for the construction of tissue specific retroviral vectors targeted towards the mammary gland. PCR primers required for the construction of such retroviral vectors have been synthesized. Retroviral vectors have been introduced into animals to determine the risk of infection of the germ line. Progeny animals have been tested for acquisition of retroviral vector sequences and new rapid protocols for the screening of large numbers of progeny animals have been established.

Tasks dealing with the stability, infectivity, and penetration of retrovi ral particles into cells, all properties dependent upon the viral glycoproteins and on the integrity of the viral envelope have been conducted. Current work uses three retroviral constructions, all derived from Moloney murine leukemia virus. Ecotropic and amphotropic producer cell lines have been prepared and the relative infectivity of the vector viruses produced are being characterized for mouse cells of different origin. Work relating to infection of cells in animals has been initiated and experimental protocols have been set up to follow the fate of virus vectors after different routes of infection.
Genetically engineered DNA molecules and viruses are being
increasingly used for biotechnological applications. Among the most widely utilized engineered molecules are retroviral vectors - viruses which have been modified to provide a highly efficient means of transporting genes into various kinds of cells. Such genetically engineered retroviruses are gaining usage in the medical field for vaccine production and gene therapy, in
research to study gene function and regulation, and in industrial applications for the production of transgenic farm-animals and bio-reactors.
The project involves (1) the establishment of both a European think-tank and a European databank on the structure and safety of retroviral vectors and on the techniques available for monitoring retroviral behaviour. The data for this databank will come from (2) a risk assessment of the use of retroviral vectors for the production of transgenic animals or for human gene therapy, and from (3) the acquisition of basic knowledge concerning the
cellular and viral mechanisms involved in modulating the behavior of retroviral vector systems. This knowledge will then be
exploited (4) to construct safe, efficient and targeted
retroviral vector systems.

Wissenschaftliches Gebiet (EuroSciVoc)

CORDIS klassifiziert Projekte mit EuroSciVoc, einer mehrsprachigen Taxonomie der Wissenschaftsbereiche, durch einen halbautomatischen Prozess, der auf Verfahren der Verarbeitung natürlicher Sprache beruht. Siehe: Das European Science Vocabulary.

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CSC - Cost-sharing contracts

Koordinator

Århus Universitet
EU-Beitrag
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Adresse
130,C.F. Møllers Allé
8000 Århus C
Dänemark

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Beteiligte (4)

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