Cytostatic process increases the productivity of cultured cells
OwnerJames E. Bailey, Martin Fussenegger +1inventors
InventorsJames E. Bailey, Martin Fussenegger, Wolfgang A. Renner
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Abstract
Provided by the invention are novel methods, vectors and cells for the recombinant production of desired gene products. In particular, the invention relates to increased production of desired gene products by inducibly arresting cell proliferation. The invention also provides novel multicistronic expression vectors that are useful not only for recombinant gene expression, but also for other applications such as gene therapy, tissue engineering and metabolic engineering.
Claims37
A method for production of a desired product from a cell, the method comprising:
- culturing a cell that has been genetically engineered to inducibly express a tumor suppressor gene product under conditions such that expression of the tumor suppressor gene product is induced, thereby arresting cell proliferation;
- and
- collecting the desired from the cultured cell.
A method for production of a desired product from a cell, the method comprising:
- culturing a cell under conditions such that expression of a tumor suppressor gene product is induced, thereby arresting cell proliferation;
- expressing an antiapoptosis gene product in the cultured cell;
- and
- collecting the desired from the cultured cell.
A method for production of a desired product from a cell, the method comprising:
- culturing a cell under conditions such that expression of a tumor suppressor gene product is induced, thereby arresting cell proliferation;
- expressing a factor that stabilizes the tumor suppressor gene product in the cultured cell;
- and
- collecting the desired from the cultured cell.
A method for production of a desired product from a cell, the method comprising:
- culturing a cell under conditions such that expression of a tumor suppressor gene product is induced, thereby arresting cell proliferation;
- expressing the product coordinately with the tumor suppressor gene product, wherein the product is selected from the group consisting of green fluorescent protein, SEAP, human growth hormone, α-interferon, β-interferon, γ-INF, insulin, erythropoietin, tissue plasminogen activator, DNAse, a monoclonal antibody, Factor VIII, Factor VII, Factor IX, HSA, IL-2, glucagon, EGF, GCSF, GMCSF, thrombopoietin, gp160 and HbSAg;
- and
- collecting the desired from the cultured cell.
A method for production of a desired product from a cell, the method comprising:
- culturing a cell under conditions such that expression of a tumor suppressor gene product is induced, thereby arresting cell proliferation;
- expressing the product coordinately with the tumor suppressor gene product, wherein the product and the tumor suppressor gene product are encoded on the same RNA;
- and
- collecting the desired from the cultured cell.
A mammalian cell that has been genetically engineered to inducibly a desired product gene and a tumor suppressor gene.
A mammalian cell that has been genetically engineered to express an antiapoptosis gene and to inducibly express a tumor suppressor gene.
A mammalian cell that has been genetically engineered to inducibly express a tumor suppressor gene and to express a factor that stabilizes the tumor suppressor gene product in the cell.
A recombinant polynucleotide vector comprising:
- a first nucleic acid sequence encoding a product protein;
- and
- a second nucleic acid sequence encoding a tumor suppressor gene product, wherein the first nucleic acid sequence is operatively linked to a first inducible promoter and the second nucleic acid sequence is operatively linked to either the first inducible promoter or to a second inducible promoter.
A recombinant polynucleotide vector comprising:
- a sequence encoding a product protein;
- a sequence encoding a tumor suppressor gene product;
- and
- an inducible promoter operatively linked to the sequence encoding a tumor suppressor gene product, wherein the vector further comprises a sequence encoding an antiapoptosis gene product or a factor that stabilizes the tumor suppressor gene product in the cell.
A host cell that contains a recombinant polynucleotide vector, wherein the host cell is selected from the group consisting of an insect cell, a fungal cell, a reptilian cell, a CHO cell, a BHK cell, a HeLa cell, a PLC cell, a Jurkat cell, a HT-1080 cell, a Hep2 cell, an ECV304 cell, a COS-7 cell, a NIH/3T3 cell, a HaCat cell, a LCL cell, a HUVEC cell, a NSO cell and a HL60 cell;
- and wherein the host cell is stably transformed by a vector comprising:
- a sequence encoding a product protein;
- a sequence encoding a tumor suppressor gene product;
- and
- an inducible promoter operatively linked to the sequence encoding the product protein and the tumor suppressor gene product.
Dependencies are inferred from the claim text.