Compositions and methods for regulating cell osmolarity
AssigneeGenentech, Inc.
InventorsLaetitia MALPHETTES, Andrew SNOWDEN, Inn H. YUK
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Abstract
The invention provides compositions and methods for regulating intracellular osmolarity in cells, e.g., in cultured cells, including in cultured cells in bioreactors. The invention provides nucleic acids comprising at least one osmo-responsive transcriptional regulatory element (OR-TRE), and cells, vectors, products of manufacture, artificial organs or implants and the like containing an osmo-responsive transcriptional regulatory element (OR-TRE).
Claims13
1-36. (canceled)
The method of claim 46 wherein the osmolality is increased by spiking with a compound that increases said osmolality.
The method of claim 46 wherein said second protein of interest is toxic to said cell.
The method of claim 46 wherein said second protein of interest is unstable.
The method of claim 46 wherein said second protein of interest is difficult to express under normal culture conditions.
41-45. (canceled)
A method for expressing a protein of interest in a cell under conditions of hyperosmolality comprising introducing a polynucleotide into a cell wherein said polynucleotide comprises:
- (a) a nucleic acid molecule comprising at least one osmo-responsive transcriptional regulatory element (OR-TRE) comprising at least one TonEBP-responsive transcriptional enhancer or NFATc-responsive transcriptional enhancer operatively linked to a transcriptional regulator and at least one Activator Protein (AP-1)-responsive transcriptional enhancer operatively linked to said TonEBP-responsive transcriptional enhancer or NFATc-responsive transcriptional enhancer and
- (b) a polynucleotide encoding a second protein of interest operatively linked to a second OR-TRE;
- wherein said nucleic acid molecule of (a) encodes TonEBP;
- and wherein under conditions of increased osmolality, said nucleic acid molecule of (a) is expressed, thereby expressing TonEBP protein, and wherein said TonEBP protein positively regulates expression of TonEBP and said second protein of interest.
52-54. (canceled)
Dependencies are inferred from the claim text.