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AssigneesMedicago Inc., Genopole
InventorsMARC-ANDRE D'AOUST, PIERRE-OLIVIER LAVOIE, ANDEOL FALCON DE LONGEVIALLE
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A method for synthesizing a protein of interest with a modified N-glycosylation profile within a plant, a portion of a plant, or a plant cell is provided. The method comprises co-expressing within a plant a nucleotide sequence encoding a first nucleotide sequence encoding a GDP-4-dehydro-6-deoxy-D-mannose reductase (RMD) the first nucleotide sequence operatively linked with a first regulatory region that is active in the plant, and a second nucleotide sequence encoding the protein of interest, the second nucleotide sequence operatively linked with a second regulatory region that is active in the plant. The first and second nucleotide sequences are co-expressed to synthesize a protein of interest comprising glycans with the modified N-glycosylation profile within the plant, the portion of the plant, or the plant cell.
A method of producing a protein of interest comprising N-glycans having a reduced fucose content in a plant, a portion of a plant, or a plant cell comprising, co-expressing within the plant, the portion of the plant, or the plant cell, a first nucleotide sequence encoding a GDP-4-dehydro-6-deoxy-D-mannose reductase (RMD), the first nucleotide sequence operatively linked with a first regulatory region that is active in the plant, the portion of a plant, or the plant cell, and a second nucleotide sequence encoding the protein of interest, the second nucleotide sequence operatively linked with a second regulatory region that is active in the plant, the portion of a plant, or the plant cell, and co-expressing the first and second nucleotide sequences to synthesize a protein of interest comprising N-glycans having a reduced fucose content, when compared to the fucose content of the protein of interest expressed in a second plant, a portion of the second plant, or a second plant cell that does not express RMD, wherein the plant, portion of the plant, or the plant cell exhibits reduced fucosylation activity resulting from both
(canceled)
A plant, a portion of a plant, or a plant cell comprising a nucleotide sequence encoding a GDP-4-dehydro-6-deoxy-D-mannose reductase (RMD), the nucleotide sequence operatively linked with a regulatory region that is active in the plant, the portion of the plant, or the plant cell, wherein the plant, the portion of the plant, or the plant cell exhibits reduced α(1,3)-fucosyltransferase (FucT) activity when compared to the FucT activity of a wildtype plant.
A method for producing a protein of interest in a plant, a portion of a plant, or a plant cell, of the Nicotiana spp having at least one of its α(1,3)-fucosyltransferase (FucT) allele knocked out comprising, co-expressing a GDP-4-dehydro-6-deoxy-D-mannose reductase (RMD) and the protein of interest within the plant, the portion of the plant, or the plant cell, to produce the protein of interest having a reduced fucosylation profile when compared to the same protein of interest produced in a second plant, a portion of the second plant, or a second plant cell having at least one of its FucT allele knocked out, and that does not express RMD, wherein the plant, the portion of the plant, or the plant cell exhibits reduced α(1,3)-fucosyltransferase (FucT) activity when compared to the FucT activity of a wildtype plant.
(canceled)
A method of producing a protein of interest comprising N-glycans having a modified N-glycosylation profile in a plant, a portion of a plant, or a plant cell comprising, co-expressing within the plant, the portion of the plant, or the plant cell, a first nucleotide sequence encoding a GDP-4-dehydro-6-deoxy-D-mannose reductase (RMD) the first nucleotide sequence operatively linked with a first regulatory region that is active in the plant, the portion of the plant, or the plant cell, and a second nucleotide sequence encoding the protein of interest, the second nucleotide sequence operatively linked with a second regulatory region that is active in the plant, the portion of the plant, or the plant cell, and co-expressing the first and second nucleotide sequences to synthesize a protein of interest comprising glycans with the modified N-glycosylation profile, when compared to the N-glycosylation profile of the protein of interest expressed in a second plant, a portion of a second plant, or a second plant cell that does not express RMD, wherein the plant, portion of the plant, or the plant cell exhibits reduced α(1,3)-fucosyltransferase (FucT) activity when compared to the FucT activity of a wildtype plant.
Dependencies are inferred from the claim text.