Abstract: Materials and methods are provided for making plants (e.g., Nicotiana varieties) that are suitable for producing therapeutic polypeptides suitable for administration to humans and animals, particularly by making TAL effector endonuclease-induced mutations in genes encoding xylosyltransferases and fucosyltransferases.

Abstract: An expression enhancer comprising a CPMV 5?UTR nucleotide sequence consisting of X nucleotides (CMPVX), where X=160, 155, 150, or 114 of SEQ ID NO:1, or consisting of a nucleotide sequence comprising from about 80% to 100% sequence similarity with CMPVX, where X=160, 155, 150, or 114 of SEQ ID NO:1 SEQ ID NO:1 is provided. The expression enhancer may further comprise a stuffer sequence fused to the 3? end of the 5?UTR nucleotide sequence (CMPVX+, where X=160, 155, 150, or 114 of SEQ ID NO:1). The stuffer sequence may comprise one or more plant kozak sequences. Plants comprising the expression enhancer and methods using the expression enhancer are also described.

Abstract: A method for synthesizing influenza virus-like particles (VLPs) within a plant or a portion of a plant is provided. The method involves expression of influenza HA in plants and the purification by size exclusion chromatography. The invention is also directed towards a VLP comprising influenza HA protein and plant lipids. The invention is also directed to a nucleic acid encoding influenza HA as well as vectors. The VLPs may be used to formulate influenza vaccines, or may be used to enrich existing vaccines.

Abstract: A method of increasing the yield, stability, or both of an acid sensitive protein in a plant is provided. The method comprises introducing a first nucleic acid and a second nucleic acid into the plant, or portion of the plant. The first nucleic acid comprises a first regulatory region active in the plant and operatively linked to a nucleotide sequence encoding the acid sensitive protein. The second nucleic acid comprises a second regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a channel protein, for example but not limited to a proton channel protein. The plant or portion of the plant is incubated under conditions that permit the expression of the nucleic acids, thereby increasing the yield of the acid sensitive protein when compared to the yield of the acid sensitive protein produced in the plant or portion of the plant produced under the same conditions, and in the absence of the proton channel protein.

Abstract: An isolated expression enhancer active in a plant, portion of a plant or plant cell, the expression enhancer is provided. The isolated expression enhancer may be selected from the group consisting of nbEPI42 (SEQ ID NO:1); nbSNS46 (SEQ ID NO:2); nbCSY65 (SEQ ID NO:3); nbHEL40 (SEQ ID NO:4); and nbSEP44 (SEQ ID NO:5). Methods for using the isolated expression enhancer are also provided.

Abstract: A method of producing a virus like particle (VLP) in a plant comprising modified H3 hemagglutinin is provided. The method comprises introducing a nucleic acid comprising a regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a modified influenza hemagglutinin (HA) protein into the plant, or portion of the plant, the modified HA protein comprises a modified proteolytic loop. Followed by incubating the plant or portion of the plant under conditions that permit the expression of the nucleic acids, thereby producing the VLP. The modified proteolytic loop may comprise one or more protease cleavage sites exhibiting reduced or abolished cleavage by a protease. Also described is a virus like particle (VLP) produced by the method, and plants expressing the VLP. The virus like particle (VLP) may comprise plant-specific N-glycans, or modified N-glycans.

Abstract: Nucleic acids encoding rotavirus VP7 fusion proteins and rotavirus-like particle (RLPs) comprising the rotavirus VP7 fusion proteins are provided. Methods for rotavirus VP7 fusion protein and RLP production in plants are also described. The VP7 fusion protein comprises a first sequence encoding a 7-1a subdomain, a second sequence encoding a 7-2 domain and a third sequence encoding a 7-1b subdomain; wherein the sequence of the 7-2 domain is derived from a first rotavirus strain and the sequence of the 7-1a subdomain, the sequence of the 7-1b subdomain or the sequence of the 7-1a subdomain and the sequence of the 7-1b subdomain are derived from a second rotavirus strain.

Abstract: The present invention relates to the production of modified influenza viral proteins in plants. More specifically, the present invention relates to producing and increasing influenza virus-like particle (VLP) production in plants, wherein the VLPs comprise the modified influenza viral proteins, such as modified influenza hemagglutinin (HA). The HA protein may comprising an amino acid sequence comprising at least one substitution when compared to a corresponding wildtype amino acid sequence. Further provided are nucleic acid encoding the modified HA protein. Furthermore methods of producing an influenza virus like particle (VLP) and methods of increasing yield of production of an influenza virus like particle (VLP) in a plant, portion of a plant, or a plant cell, are also provided.

Abstract: The present invention relates to the production of modified influenza vial proteins in plants. More specifically, the present invention relates to producing and increasing influenza virus-like particle (VLP) production in plants, wherein the VLPs comprise the modified influenza viral proteins, such as modified influenza hemagglutinin (HA). The H A protein may comprising an amino acid sequence comprising at least one substitution when compared to a corresponding wildtype amino acid sequence. Further provided are nucleic acid encoding the modified H A protein. Furthermore methods of producing an influenza virus like particle (VLP) and methods of increasing yield of production of an influenza virus like particle (VLP) in a plant, portion of a plant, or a plant cell, are also provided.

Abstract: A method of producing a virus like particle (VLP) in a plant is provided. The method comprises introducing a first nucleic acid and a second nucleic acid into the plant, or portion of the plant. The first nucleic acid comprises a first regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a structural virus protein. The second nucleic acid comprises a second regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a channel protein, for example but not limited to a proton channel protein. The plant or portion of the plant is incubated under conditions that permit the expression of the nucleic acids, thereby producing the VLP.

Abstract: An isolated expression enhancer active in a plant, portion of a plant or plant cell, the expression enhancer is provided. The isolated expression enhancer may be selected from the group consisting of nbMT78 (SEQ ID NO:1); nbATL75 (SEQ ID NO:2); nbDJ46 (SEQ ID NO:3); nbCHP79 (SEQ ID NO:4); nbEN42 (SEQ ID NO:5); atHSP69 (SEQ ID NO:6); atGRP62 (SEQ ID NO:7); atPK65 (SEQ ID NO:8); atRP46 (SEQ ID NO:9); nb30S72 (SEQ ID NO:10); nbGT61 (SEQ ID NO:11); nbPV55 (SEQ ID NO:12); nbPPI43 (SEQ ID NO:13); nbPM64 (SEQ ID NO:14); and nbH2A86 (SEQ ID NO:15). Methods for using the isolated expression enhancer are also provided.

Abstract: A method of producing a virus like particle (VLP) in a plant comprising modified hemagglutinin is provided. The method comprises introducing a nucleic acid comprising a regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a modified influenza hemagglutinin (HA) protein into the plant, or portion of the plant, the modified HA protein comprises a modified proteolytic loop. Followed by incubating the plant or portion of the plant under conditions that permit the expression of the nucleic acids, thereby producing the VLP. The modified proteolytic loop may comprise one or more protease cleavage sites exhibiting reduced or abolished cleavage by a protease. The nucleotide sequence encoding the HA may be selected from the group consisting of B HA, C, H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, and H16. Also described is a virus like particle (VLP) produced by the method, and plants expressing the VLP.

Abstract: The present invention relates to the production of modified influenza viral proteins in plants. More specifically, the present invention relates to producing and increasing influenza virus-like particle (VLP) production in plants, wherein the VLPs comprise the modified influenza viral proteins, such as modified influenza hemagglutinin (HA). The HA protein may comprising an amino acid sequence comprising at least one substitution when compared to a corresponding wildtype amino acid sequence. Further provided are nucleic acid encoding the modified HA protein. Furthermore methods of producing an influenza virus like particle (VLP) and methods of increasing yield of production of an influenza virus like particle (VLP) in a plant, portion of a plant, or a plant cell, are also provided.

Abstract: Nucleic acids encoding modified norovirus VP1 proteins, and VLPs comprising one or more of the modified norovirus VP1 proteins are provided. Methods for modified norovirus VP1 protein, and norovirus VLP, production in plants, portions of the plant or a plant cell, are also described.

Abstract: Materials and methods are provided for making plants (e.g., Nicotiana varieties) that are suitable for producing therapeutic polypeptides suitable for administration to humans and animals, particularly by making TAL effector endonuclease-induced mutations in genes encoding xylosyltransferases and fucosyltransferases.

Abstract: A method of increasing expression of an heterologous protein of interest in a plant or portion of the plant is provided. The method comprises treating the plant or portion of the plant with a jasmonate-pathway activator, and introducing a nucleotide sequence operably linked to a regulatory region derived from a DNA plant virus and encoding the heterologous protein of interest into the plant or portion of the plant. Alternatively, the plant or plant portion may comprise the nucleic acid and encoding the heterologous protein of interest, and the plant or portion of the plant is treated with the jasmonate pathway activator. The treated plant is incubated under conditions to permit expression of the nucleotide sequence encoding the heterologous protein of interest.

Abstract: A method of producing a virus like particle (VLP) in a plant, and compositions comprising VLPs, are provided. The method involves introducing a nucleic acid comprising a regulatory region active in the plant and operatively linked to a chimeric nucleotide sequence encoding, in series, an ectodomain from a virus trimeric surface protein or fragment thereof, fused to an influenza transmembrane domain and cytoplasmic tail, into the plant, or portion of the plant, the ectodomain is from a non-influenza virus trimeric surface protein and heterologous with respect to the influenza transmembrane domain, and the cytoplasmic tail. The plant or portion of the plant are incubated under conditions that permit the expression of the nucleic acid, thereby producing the VLP. A VLP produced by this method are also provided.

Abstract: A method of producing a virus like particle (VLP) in a plant, and compositions comprising VLPs, are provided. The method involves introducing a nucleic acid comprising a regulatory region active in the plant and operatively linked to a chimeric nucleotide sequence encoding, in series, an ectodomain from a virus trimeric surface protein or fragment thereof, fused to an influenza transmembrane domain and cytoplasmic tail, into the plant, or portion of the plant, the ectodomain is from a non-influenza virus trimeric surface protein and heterologous with respect to the influenza transmembrane domain, and the cytoplasmic tail. The plant or portion of the plant are incubated under conditions that permit the expression of the nucleic acid, thereby producing the VLP. A VLP produced by this method are also provided.

Abstract: Nucleic acids encoding modified norovirus VP1 proteins, and VLPs comprising one or more of the modified norovirus VP1 proteins are provided. Methods for modified norovirus VP1 protein, and norovirus VLP, production in plants, portions of the plant or a plant cell, are also described.

Abstract: An expression enhancer comprising a CPMV 5?UTR nucleotide sequence consisting of X nucleotides (CMPVX), where X=160, 155, 150, or 114 of SEQ ID NO:1, or consisting of a nucleotide sequence comprising from about 80% to 100% sequence similarity with CMPVX, where X=160, 155, 150, or 114 of SEQ ID NO:1 SEQ ID NO:1 is provided. The expression enhancer may further comprise a stuffer sequence fused to the 3? end of the 5?UTR nucleotide sequence (CMPVX+, where X=160, 155, 150, or 114 of SEQ ID NO:1). The stuffer sequence may comprise one or more plant kozak sequences.