Viral vector production system

A viral vector production system comprising:

• 1) a viral vector production cell comprising nucleic acid sequences encoding viral vector components;
• and 2) a nuclease helper cell comprising a nucleic acid sequence encoding a nuclease, wherein the nuclease is expressed and secreted in co-culture of the production cell of 1) and the helper cell of 2), thereby degrading residual nucleic acid during viral vector production.

A method of producing a viral vector, the method comprising, transfecting a viral vector production cell with nucleic acid sequences encoding:

• 1) viral vector components;
• and 2) a nuclease, wherein the viral vector components and the nuclease are expressed in the viral vector production cell and secreted in cell culture thereby degrading residual nucleic acid during viral vector production.

A method of producing a viral vector, the method comprising contacting 1) a viral vector production cell expressing viral vector components with 2) a nuclease helper cell expressing a nuclease, wherein the nuclease is expressed in the helper cell and secreted in co-culture of the production cell with the helper cell thereby degrading residual nucleic acid during viral vector production.

A method of producing a viral vector, the method comprising contacting 1) a viral vector production cell expressing viral vector components with 2) a liquid feed from nuclease helper cell expressing a nuclease, wherein the nuclease is expressed in the cell and secreted in cell culture thereby degrading residual nucleic acid during viral vector production.

In an improved method of producing a viral vector, the improvement comprising introducing nucleic acid sequences into a viral vector production cell, wherein the nucleic acid sequences encode:

• 1) viral vector components;
• and 2) a nuclease, wherein the nuclease is expressed in the production cell and secreted in cell culture thereby degrading residual nucleic acid during viral vector production.

In an improved method of producing a viral vector, the improvement comprising contacting in co-culture a viral vector production cell expressing viral vector components with a nuclease helper cell expressing a nuclease, wherein the nuclease is expressed in the production cell and secreted in cell culture thereby degrading residual nucleic acid during viral vector production.

(canceled)

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11-13. (canceled)

15-31. (canceled)

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36-41. (canceled)

A production cell for producing viral vectors comprising nucleic acid sequences encoding:

• 1) viral vector components;
• and 2) a nuclease, wherein the nuclease is expressed in the viral vector production cell and secreted in cell culture thereby degrading residual nucleic acid during viral vector production, and wherein the production cell is a eukaryotic production cell.

45-51. (canceled)

A production cell for producing viral vectors comprising nucleic acid sequences encoding:

• 1) viral vector components;
• and 2) a nuclease fusion protein, wherein the nuclease fusion protein comprises an exonuclease domain fused to an endonuclease domain, and wherein the nuclease fusion protein is expressed in the viral vector production cell and secreted in cell culture thereby degrading residual nucleic acid during viral vector production, and wherein the production cell is a eukaryotic production cell.

(canceled)

A cell culture device comprising a viral vector production system comprising a viral vector production cell comprising nucleic acid sequences encoding:

• 1) viral vector components;
• and 2) a nuclease, wherein the nuclease is expressed in the production cell and secreted in cell culture thereby degrading residual nucleic acid during viral vector production.

(canceled)

A variant of a secreted nuclease capable of degrading residual nucleic acid during viral vector production, said variant comprising the amino acid sequence of SEQ ID NO:

• 11.

A modified nuclease having increased cell-retention and/or or cell-association that is expressed through the secretory pathway of a eukaryotic cell, wherein the modified nuclease comprises a retention signal at its C-terminus.

59-71. (canceled)

73-76. (canceled)

A nuclease helper cell wherein secreted nuclease activity within the helper cell culture is at least about 10 unit per mL of equivalent Benzonase® nuclease activity as determinable by the assay presented as Assay 1 herein.

78-82. (canceled)