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 virus-like particles (VLPs) containing one or more of the modified norovirus VP1 proteins, and compositions containing the nucleic acids, VP1 proteins, or the VLPs are provided. Methods for producing modified norovirus VP1 protein, and norovirus VLP, production in plants, portions of the plant or a plant cell, are also provided.

Abstract: Modified coronaviras spike (S)-protein, virus-like particle (VLPs) comprising the modified S protein and nucleic acids encoding modified S protein are provided. Methods for modified S-protein and VLP production in a host or host cell are also described. The modified S-protein may comprise a transmembrane domain (TM) or portion of a TM, and a cytosolic tail (CT) or of a CT, wherein the CT or portion of the CT is derived from an influenza hemagglutinin (HA) protein and wherein the TM or portion of the TM is heterologous to the CT or portion of the CT. In addition a method for inducing immunity to a coronaviras infection in a subject, comprising administering a composition comprising the modified coronaviras (S)-protein or VLP to the subject is described.

Abstract: Nucleic acids encoding norovirus VP1 fusion proteins and VLPs comprising the norovirus VP1 fusion proteins are provided. Methods for norovirus VP1 fusion protein and norovirus VLP production in plants are also described. The VP1 fusion protein comprises, a first sequence encoding an S domain derived from a first norovirus strain, and a second sequence encoding a P domain derived from a second norovirus strain.

Abstract: A method for synthesizing a protein of interest within a plant or a portion of a plant is provided. The method involves treating the plant to increase secondary leaf biomass production, followed by introducing one or more than one nucleic acid sequence encoding a protein of interest operatively linked with a regulatory region that is active in the plant, into the plant. The plant is then maintained under conditions that permit the nucleic acid sequence encoding the protein of interest to be expressed in the plant or the portion of the plant. Optionally, the plant or portion of the plant may be harvested and the protein of interest extracted.

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.

Abstract: Methods of preparing plant-derived proteins or suprastructure proteins, are provided. The method may comprise obtaining a plant, or plant matter comprising apoplast-localized proteins, or suprastructure proteins, producing a protoplast/spheroplast fraction and apoplast fraction from the plant or plant matter, and recovering the apoplast fraction. The apoplast fraction comprises plant-derived proteins or suprastructure proteins. Alternatively, the proteins, or suprastructure proteins, may be obtained from plant or plant matter comprising plant-derived proteins or suprastructure proteins, by digesting the plant matter using a cell wall degrading enzyme composition to produced a digested fraction. The digested fraction is filtered to produced a filtered fraction, and the plant-derived proteins or suprastructure proteins, are recovered from the filtered fraction.

Abstract: Methods of preparing plant-derived virus like particles (VLPs) are provided. The method may comprise obtaining a plant, or plant matter comprising apoplast-localized VLPs, producing a protoplast/spheroplast fraction and apoplast fraction from the plant or plant matter, and recovering the apoplast fraction. The apoplast fraction comprises plant-derived VLPs. Alternatively, VLPs may be obtained from plant or plant matter comprising plant-derived VLPs by digesting the plant matter using a cell wall degrading enzyme composition to produced a digested fraction. The digested fraction is filtered to produced a filtered fraction, and the plant-derived VLPs are recovered from the filtered fraction.

Abstract: Nucleic acids encoding modified norovirus VP1 proteins, and virus-like particles (VLPs) containing one or more of the modified norovirus VP1 proteins, and compositions containing the nucleic acids, VP1 proteins, or the VLPs are provided. Methods for producing modified norovirus VP1 protein, and norovirus VLP, production in plants, portions of the 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: A suprastructure comprising a modified influenza hemagglutinin (HA) is provided. The modified HA may comprise one or more than one alteration that reduces non-cognate binding of the modified HA to sialic acid (SA) on the surface of a cell, while maintaining cognate interaction with the cell, such as a B cell. A composition comprising the suprastructure and modified HA and a pharmaceutically acceptable carrier is also described. A method of increasing an immunological response or inducing immunity in response to a vaccine comprising the suprastructure and modified HA is also provided.

Abstract: Nucleic acids encoding norovirus VP1 fusion proteins and VLPs comprising the norovirus VP1 fusion proteins are provided. Methods for norovirus VP1 fusion protein and norovirus VLP production in plants are also described. The VP1 fusion protein comprises, a first sequence encoding an S domain derived from a first norovirus strain, and a second sequence encoding a P domain derived from a second norovirus strain.

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 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: Nucleic acids encoding norovirus VP1 fusion proteins and VLPs comprising the norovirus VP1 fusion proteins are provided. Methods for norovirus VP1 fusion protein and norovirus VLP production in plants are also described. The VP1 fusion protein comprises, a first sequence encoding an S domain derived from a first norovirus strain, and a second sequence encoding a P domain derived from a second norovirus strain.

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.