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AssigneeGesellschaft fuer Biotechnologische Forschung mbH (GBF)
InventorsDagmar Wirth, Dirk Spitzer, Hansjoerg Hauser
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Murine retroviruses are the most important transfer systems for human gene therapy. However, their application is currently limited. One of the major restrictions both for an application in vivo resides in the problem that this virus type is sensitive to inactivation by human complement factors. Our invention overcomes this limitation. We have modified murine recombinant retroviruses in a way that they are resistant to human complement factors. This was achieved by genetic modification of the retroviral surface protein env which is responsible for receptor interaction: the receptor interacting domain of env was fused to catalytically active domains of human complement inactivation factors. These modified env were expressed in complement-sensitive cells and specifically integrated into virus particles. By this strategy cells and viruses are generated that are fully resistant to complement attack. Thus, this strategy provides a tool for establishment of complement resistant cells and generation of viruses for in vivo gene therapy.
Single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) encoding a fusion protein of
A method of producing a virus stable against a human complement system for the enhancement of the transduction efficiency of the virus for gene therapy by fusing a retroviral surface protein (env) or part thereof comprising its transmembrane domain to the catalytically active domains of a complement inactivation factor.
A method of producing a fusion protein by fusing a retroviral surface protein (env) or part thereof comprising its transmembrane domain to the catalytically active domain of a complement inactivation factor for the generation of complement resistant cells for transplantation.
A method of producing a fusion protein by fusing a retroviral surface protein (env) or part thereof comprising its transmembrane domain to the catalytically active domain of a complement inactivation factor for the reduction of the risk of the formation of replication competent viruses by means of packaging cells via recombination by eliminating the intrinsic sequence overlap of retroviral gagpol and env genes.
A method of producing a fusion protein by fusing a retroviral surface protein (env) or part thereof comprising its transmembrane domain to the catalytically active domain of a complement inactivation factor for generating complement stabilized packaging cells by coexpression of wild-type env proteins and fusion proteins.
A method of producing a fusion protein by fusing a retroviral surface protein (env) or part thereof comprising its transmembrane domain to the catalytically active domain of a complement inactivation factor for generating complement stabilized viruses.
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