Herpes simplex virus strains for gene transfer
AssigneeUniversity of Pittsburgh—Of the Commonwealth System of Higher Education
InventorNeal A. DeLuca
Life of this patent
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Abstract
Cell lines that express complementing levels of herpes simplex virus (HSV) essential immediate early proteins ICP4 and ICP27 as well as ICP4, ICP27 and ICP0 and a method of producing the novel cell lines are disclosed. These cell lines are utilized to provide HSV strains deficient for both (a) ICP4 and ICP27; (b) ICP4, ICP27, ICP22; (c) ICP4, ICP27, ICP0; and, (d) ICP4, ICP27, ICP22 and ICP0, and their generation, and HSV strains deficient for (a) ICP4 and ICP27; (b) ICP4, ICP27, ICP22; (c) ICP4, ICP27, ICP0; and, (d) ICP4, ICP27, ICP22 and ICP0, and one or more additional genes, and their generation. Vectors are provided from these methods of using these HSV strains for gene transfer and for producing site-specific homologous recombination with cellular DNA.
Claims47
A cell line which comprises DNA encoding herpes simplex virus (HSV) proteins ICP4, ICP27, and ICP0.
An HSV strain comprising a genome defective for the HSV ICP4, ICP27, and ICP0 genes.
An HSV vector comprising
- (a) a genome defective for the HSV ICP4, ICP27, and ICP0 genes, and
- (b) at least one exogenous gene.
A method of producing a mutant HSV which comprises:
- infecting an ICP4.sup.(+) /ICP27.sup.(+) /ICP0.sup.(+) cell line expressing complementing levels of ICP4, ICP27, and ICP0 with a mutant HSV;
- incubating said cell line in a tissue culture medium;
- and,
- collecting said mutant HSV produced by said cell line.
A method of expressing an exogenous gene within an isolated mammalian cell which comprises infecting said cell with an HSV vector comprising a genome (a) defective for the HSV ICP4, ICP27, and ICP0 genes, and (b) comprising said exogenous gene, whereby said gene is expressed within said cell.
A method of directing homologous recombination with cellular DNA, said method comprising infecting an isolated cell with an HSV vector, said vector comprising a genome (a) defective for the HSV ICP4 and ICP27 genes and (b) comprising a DNA sequence homologous to said cellular DNA, whereby homologous recombination occurs between said genome and said cellular DNA.
Dependencies are inferred from the claim text.