Provided herein are Vero cell lines that stably express Herpes Simplex Virus (HSV) ICP0 protein. These cells have the same morphology of Vero cells, exhibit stable expression of HSV ICP0 protein, and also efficiently complement replication of HSV ICP0 deficient virus for greater than 20, 30, or even 40 cell passages.

The present invention is directed to a recombinant herpesvirus which comprises the GCN4 yeast transcription factor or a part thereof fused to or inserted into glycoprotein H and is capable of binding to a target molecule present on a cell for propagation and production of the herpesvirus. The herpesvirus may comprise additional modification in glycoprotein D and/or glycoprotein B for retargeting the herpesvirus to a diseased cell. The present invention is further directed to a nucleic acid and a vector coding for the gH, a polypeptide comprising the gH, and a cell comprising the herpesvirus, nucleic acid, vector or polypeptide. Moreover, the present invention is directed to a cell having accessible on the surface a target molecule for the GCN4 yeast transcription factor or part thereof and to a method for producing the herpesvirus in said cell.

In one embodiment, the invention provides a herpes simplex virus (HSV) vector that does not express toxic HSV genes in non-complementing cells and which comprises a genome comprising one or more transgenes, wherein the vector is capable of expression of a transgene for at least 28 days in non-complementing cells. The invention also relates to viral stocks of the inventive vectors, compositions thereof suitable for use therapeutically or for in vitro applications, and methods relating thereto. In another aspect, the invention provides a complementing cell engineered to express ICP4 and ICP27 when the cell is infected with HSV for the production of the inventive vector.

A genetically modified mammalian cell and genetically modified mammalian cell line comprise a recombination sequence inserted in a target locus on a chromosome of the mammalian cell genome, wherein the recombination sequence comprises Bxb1attB sequence from Mycobacterium smegmatis. A transgenic mammalian cell and transgenic mammalian cell line comprise a heterologous nucleic acid stably integrated in a target locus on a chromosome of the mammalian genome, wherein the heterologous nucleic comprises a heterologous gene configured for expression by the transgenic mammalian cell.

Methods and compositions for increasing the production of recombinant proteins by introducing ICP0 to cells capable of producing a recombinant protein are encompassed. In one method, the recombinant protein is a protein that is required for the replication of a replication defective virus, wherein the recombinant protein is provided to the replication defective virus in trans.

Methods and compositions for increasing the production of recombinant proteins by introducing ICP0 to cells capable of producing a recombinant protein are encompassed. In one method, the recombinant protein is a protein that is required for the replication of a replication defective virus, wherein the recombinant protein is provided to the replication defective virus in trans.

Malignant tumors that are resistant to conventional therapies represent significant therapeutic challenges. An embodiment of the present invention provides an oncolytic virus capable of killing target cells, such as tumor cells. In various embodiments presented herein, the oncolytic viruses described herein are suitable for treatment of several types of cancer, including glioblastoma.

Disclosed are custom HSV-1 packaging genomes for chromosomal integration into a cell line that is devoid of cis-acting HSV origins of replication (ori) and packaging sequences (pac) normally used for incorporation of the viral genome into virus particles, as well as the use of the resulting packaging cell for amplicon packaging.

The invention described herein provides a recombinant replication-defective virus derived from Herpesvirales order, comprising a defective ICP27 gene that impairs or otherwise does not support replication of the HSV, or a functional equivalent gene thereof; a gene-of-interest (GOI) flanked by AAV ITR sequences inserted at, within, or replacing a non-essential or replaceable essential locus of the HSV; and a coding sequence for AAV Rep and Cap proteins inserted at, within or replacing the non-essential or replaceable essential locus of the HSV. The invention also provides production cell lines for such recombinant replication-defective virus, wherein the cell lines have a coding sequence for ICP27 or a functional equivalent thereof, and wherein the coding sequence has no or minimal sequence overlap with the virus characterized by the defective ICP27 gene that impairs or otherwise does not support replication of the HSV. Methods of using such recombinant replication-defective virus and production cell lines are also provided. rAAV produced using such recombinant replication-defective virus and/or production cell lines and methods of making such rAAV are also provided.

Methods and compositions for increasing the production of recombinant proteins by introducing ICP0 to cells capable of producing a recombinant protein are encompassed. In one method, the recombinant protein is a protein that is required for the replication of a replication defective virus, wherein the recombinant protein is provided to the replication defective virus in trans.