Recombinant viruses displaying a nonviral polypeptide on their e

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage

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435 6, 4352351, 536 234, 536 2372, C12Q 170, C12Q 168, C12N 100, C07H 2104

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057232873

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BRIEF SUMMARY
FIELD OF INVENTION

This invention relates to recombinant viruses, also referred to as recombinant viral particles. By "recombinant virus", we mean a virus in which at least one of the components of the virion particle is altered or derived by recombinant DNA technology.
This invention also relates to the field of therapeutic gene transfer and concerns the teleological design and use of vectors to derive recombinant proteins or protein components suitable for display on the surface of a gene delivery vehicle which, when displayed on the surface of the gene delivery vehicle, through their interaction with components of the surface of a eukaryotic target cell, are capable of influencing the efficiency with which the gene delivery vehicle delivers its nucleic acid into the target cell, or of transmitting a signal to the target cell which influences the subsequent fate of the delivered nucleic acid, and which are thereby capable of enhancing the suitability of the, gene delivery vehicle for an intended application.


BACKGROUND TO THE INVENTION
eukaryotic cells
Recombinant viruses have been widely used as vectors for the delivery of foreign genes into eukaryotic cells. Recombinant viruses which are used for delivery of foreign genes to animal cells include members of several virus families, including Adenovirus, Herpesvirus, Togavirus, and Retrovirus families. Viruses which infect eukaryotic cells comprise a protein shell or shells (the capsid) formed by the multimeric assembly of multiple copies of one or more virus-encoded proteins. The capsid houses the viral nucleic acid (RNA or DNA) and may or may not be enveloped in a lipid bilayer which is studded with virus-encoded oligomeric spike glycoproteins, visible on electron micrographs as spikes projecting from the surface of the virus.
The initial event in the virus life cycle is binding to the surface of the eukaryotic target cell. Binding is mediated by the direct interaction of specialised proteins or glycoproteins on the surface of the virus (antireceptors) with receptors on the surface of the target cell, or indirectly via soluble ligands which bind the virus to receptors on the surface of the target cell. In some instances, the interaction between a virus and a target cell receptor may transmit a metabolic signal to the interior of the target cell. Binding is followed by penetration of the target cell membrane and entry of the viral nucleic acid into the cytosol (reviewed in Marsh and Helenius 1989 Adv Virus Res 36 p 107-151). Some nonenveloped viruses undergo conformational changes which result in their direct translocation across the target cell membrane, whereas others, such as adenovirus, are first endocytosed and then cause disruption of the wall of the acidified endosomal vesicle. Enveloped viruses fuse with the target cell plasma membrane whereupon the virus capsid (or core particle), housing the viral nucleic acid is released into the cytoplasm of the target cell. This envelope fusion event is catalysed by oligomeric viral membrane spike glycoproteins which are anchored in the viral envelope and may, or may not be dependent on the prior endocytosis of bound virus and its exposure to an acidic environment within the endosomal vesicle. The mechanisms by which viral spike glycoproteins catalyse membrane fusion may involve their proteolytic cleavage, the dissociation of noncovalently linked subunits or other conformational alterations which expose buried hydrophobic moieties capable of penetrating the lipid membrane of the target cell. Thus, virus-mediated delivery of nucleic acid is a complex, multistage process.
After delivery of the viral nucleic acid into the target cell, further steps in the viral life cycle which lead to viral gene expression, genome replication and the production of progeny viruses are often critically dependent on variable host cell factors. For example, division of the infected target cell is required for efficient integration of a reverse-transcribed retroviral genome into the host cell chromosome and subsequent retrovira

REFERENCES:
London et al: "Infectious enveloped RNA virus antigenis chimeras", Proc.Natl.Acad. Sci. USA, vol. 89, 1992 pp. 207-211, see the whole document.
Cesareni; "Peptide display on filamentous phage capsides", FEBS Letters, vol. 307, No. 1, 1992, pp. 66-70, see the whole document.
Dong et al.: "A chimeric avian retrovirus containing the influenza virus hemagglutinen gene", J. Virol. vol. 66, No. 12, 1992, pp. 7374-7382, see p. 7381, col. 2, line 40-line 53.

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