Inducible promoter for viral vector production
AssigneeAsklepios Biopharmaceutical, Inc.
InventorsGraham Whyteside, Michael L. Roberts, Victoria Fiona Torrance
Life of this patent
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Abstract
Aspects described herein relate to stable cell line for recombinant viral vector (e.g., recombinant adeno associated viral vector) production comprising at least one inducible promoter operatively linked to a heterologous gene that encodes a toxic protein. Methods for making stable cell lines, and methods for viral vector production are further provided herein.
Claims29
A nucleic acid construct for use in production of viral vectors, comprising:
- a nucleic acid sequence comprising at least one of:
- a nucleic acid sequence encoding a E4 protein, a nucleic acid sequence encoding a E2A protein, and a nucleic acid sequence encoding a VA RNA, wherein each nucleic acid sequences encoding any one of E4 protein, E2A protein, and VA RNA is operatively linked to a first regulatable promoter;
- and
- a nucleic acid sequence encoding a Rep protein, wherein the nucleic acid encoding a Rep protein is under the control of a second regulatable promoter or heterologous transcriptional activator,
- wherein the first and the second regulatable promoters are different,
- wherein at least the first or second regulatable promoter is selected from the group consisting of a forskolin inducible promoter selected from SEQ ID NO:
- 4-6 or a variant thereof, and a hypoxia inducible promoter selected from SEQ ID NO:
- 1-3, or a variant thereof.
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A nucleic acid construct comprising
- a first nucleic acid construct comprising in a 5′ to 3′ direction:
- a promoter, a stop nucleic acid sequence flanked by a first pair of recombinase recognition sequences (RRS), and nucleic acid sequence encoding a Rep protein, wherein the promoter is operatively linked to the nucleic acid encoding the Rep protein, and
- a second nucleic acid construct comprising, in a 5′ to 3′ direction:
- a promoter, a stop nucleic acid sequence flanked by a second pair of recombinase recognition sequences (RRSs), and nucleic acid sequence encoding one or more of E2A, E4, and VA RNA, wherein the promoter is operatively linked to the nucleic acid encoding the one or more of E2A, E4, and VA RNA,
- wherein the promoter is selected from the group consisting of a forskolin inducible promoter selected from SEQ ID NO:
- 4-6 or a variant thereof, and a hypoxia inducible promoter selected from SEQ ID NO:
- 1-3, or a variant thereof.
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Dependencies are inferred from the claim text.