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Last Updated: March 29, 2024

Claims for Patent: 7,781,197


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Summary for Patent: 7,781,197
Title:Transformed bryophyte cell having disrupted endogenous alpha 1,3-fucosyl and beta 1,2-xylosyl transferase encoding nucleotide sequences for the production of heterologous glycosylated proteins
Abstract: Bryophyte plants and bryophyte plant cells comprising dysfunctional fucT and xylT genes and an introduced glycosyltransferase gene, methods for the production of glycosylated proteins therewith, vectors and uses thereof.
Inventor(s): Reski; Ralf (Oberried-Zastler, DE), Gorr; Gilbert (Freiburg, DE), Decker; Eva (Freiburg, DE), Stemmer; Christian (Freiburg, DE), Lienhart; Otmar (Freiburg, DE), Koprivova; Anna (Freiburg, DE)
Assignee: greenovation Biotech GmbH (Freiburg, DE)
Application Number:10/539,725
Patent Claims:1. A transformed bryophyte cell from Physcomitrella patens that comprises i) a disrupted endogenous alpha 1,3-fucosyl transferase encoding nucleotide sequence and ii) a disrupted endogenous beta 1,2-xylosyl transferase encoding nucleotide sequence, whereby the bryophyte cell is incapable of forming N-linked glycans with 1,3-linked fucosyl and 1,2-linked xylosyl residues.

2. A transformed bryophyte cell according to claim 1, wherein the cell further comprises a nucleotide sequence operably linked to an exogenous promoter that drives expression in the bryophyte cell, wherein said nucleotide sequence encodes a glycosylated polypeptide that is expressed in the bryophyte cell.

3. A transformed bryophyte cell according to claim 1, further comprising a nucleotide sequence operably linked to an exogenous promoter that drives expression in the bryophyte cell, wherein said nucleotide sequence encodes a functional human beta 1, 4 galactosyltransferase that is expressed in the bryophyte cell, whereby the bryophyte cell is capable of forming N-linked glycans with terminal 1,4-linked galactosyl residues.

4. A transformed bryophyte cell according to claim 2, wherein said glycosylated polypeptide is a polypeptide having a primary amino acid sequence of a human glycosylated polypeptide or a primary amino acid sequence of an antibody or an active fragment thereof.

5. A transformed bryophyte cell according to claim 4, wherein said glycosylated polypeptide is selected from the group consisting of human insulin, preproinsulin, vascular endothelial growth factor (VEGF), proinsulin, glucagon, alpha-interferon, beta-interferon, gamma-interferon, blood-clotting factors VII, VIII, IX, X, XI, and XII, luteinising hormone, follicle stimulating hormone, epidermal growth factor, platelet-derived growth factor, granulocyte colony stimulating factor, prolactin, oxytocin, thyroid stimulating hormone, adrenocorticotropic hormone, calcitonin, parathyroid hormone, somatostatin, erythropoietin (EPO), beta-glucocerebrosidase, haemoglobin, serum albumin, and collagen.

6. A bryophyte plant or bryophyte tissue from Physcomitrella patens comprising a transformed bryophyte cell according to claim 1.

7. A transformed bryophyte cell according to claim 1, wherein said disrupted alpha 1,3-fucosyl transferase encoding nucleotide sequence and said disrupted beta 1,2-xylosyl transferase encoding nucleotide sequence are each independently disrupted by insertion of exogenous nucleic acids or by at least partial deletion of endogenous nucleic acids.

8. A transformed bryophyte cell according to claim 4, wherein said glycosylated polypeptide is selected from the group consisting of an interferon, a fertility hormone, a growth factor and an enzyme.

9. A transformed bryophyte cell according to claim 2, further comprising a nucleotide sequence operably linked to an exogenous promoter that drives expression in the bryophyte cell, wherein said nucleotide sequence encodes a functional human beta 1, 4 galactosyltransferase that is expressed in the bryophyte cell, whereby the bryophyte cell is capable of forming N-linked glycans with terminal 1,4-linked galactosyl residues.

10. A transformed bryophyte cell according to claim 9, wherein said glycosylated polypeptide is a polypeptide having a primary amino acid sequence of a human glycosylated polypeptide or a primary amino acid sequence of an antibody or an active fragment thereof.

11. A transformed bryophyte cell according to claim 10, wherein said glycosylated polypeptide is selected from the group consisting of human insulin, preproinsulin, vascular endothelial growth factor (VEGF), proinsulin, glucagon, alpha-interferon, beta-interferon, gamma-interferon, blood-clotting factors VII, VIII, IX, X, XI, and XII, luteinising hormone, follicle stimulating hormone, epidermal growth factor, platelet-derived growth factor, granulocyte colony stimulating factor, prolactin, oxytocin, thyroid stimulating hormone, adrenocorticotropic hormone, calcitonin, parathyroid hormone, somatostatin, erythropoietin (EPO), beta-glucocerebrosidase, haemoglobin, serum albumin, and collagen.

12. A bryophyte plant or bryophyte tissue from Physcomitrella patens comprising a transformed bryophyte cell according to claim 9.

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