The present invention relates to methods and compositions for enhancing expression from RNA expression vectores. The invention is based upon the observation that reducing the frequency of the dinucleotide CpG and UpA has a significant effect on expression from such vectores. Aspects of the invention include, amongst others, synthetic RNA vectores, virions, cells, methods of producing vaccines and methods of treatment or immunisation.

Disclosed herein are methods and exemplary compositions associated with virus purification, antigen purification, and conjugation of virus and proteins (e.g., antigen) to form vaccines for delivery of immunological and other therapeutic agents, exemplary aspects of which may include harvesting viral and antigenic substances from source organisms; a purification platform comprising chemical separation and size-difference separation for the removal of contaminants, debris and impurities from the viral and protein (e.g. antigenic, including influenza hemagglutinin antigens) substances, as well as their concentration and collection; and a conjugation platform providing activation of the virus at a pH that increases binding rate and binding propensity between the virus and the protein, wherein embodiments related to the conjugation platform include controlling the ratio of virus to protein.

The present invention is directed to improved methods for conjugating polyethylene glycol to polyethyleneimine, and to the use of such polyethylene glycol—polyethyleneimine conjugates to improve the efficiency with which viral and non-viral nucleic acid vectors transfect cells to provide gene therapy, and to improve the efficiency of producing viral particles that comprise therapeutic polynucleotides for use in gene therapy.

Methods and compositions for selecting plants with targeted nuclease alterations are provided.

A Barley stripe mosaic virus-based gene editing vector system, comprising artificial plasmids separately containing Barley stripe mosaic virus RNAα, RNAβ, and RNAγ. The required sgRNA sequence is integrated in RNAβ or RNAγ. The Barley stripe mosaic virus-based gene editing vector system can perform efficient gene editing on genomes of dicotyledons such as Nicotiana benthamiana and monocotyledons such as wheat and maize. Using the gene editing vector system, users can directly obtain wheat seeds harboring targeted gene editing events simply by inoculating the Cas9-transgenic wheat, and the edited target gene is transmitted to progeny plants through the seeds, without the need for transformation, tissue culture and regeneration process

The present disclosure relates to a single stranded RNA vector suitable for introducing a therapeutic agent, such as a peptide, a protein or a small RNA, into a host plant. The vector does not encode for any movement protein or coat protein, but is capable of capable of systemic and phloem-limited movement and replication within the host plant.

Disclosed herein are methods and exemplary compositions associated with antigen purification, exemplary aspects of which may include harvesting viral and antigenic substances from source organisms; and a purification platform comprising chemical separation and size-difference separation for the removal of contaminants, debris and impurities from the viral and protein (e.g. antigenic, including influenza hemagglutinin antigens) substances, as well as their concentration and collection.

Disclosed herein are methods and exemplary compositions associated with virus purification, antigen purification, and conjugation of virus and proteins (e.g., antigen) to form vaccines for delivery of immunological and other therapeutic agents, exemplary aspects of which may include harvesting viral and antigenic substances from source organisms; a purification platform comprising chemical separation and size-difference separation for the removal of contaminants, debris and impurities from the viral and protein (e.g. antigenic, including influenza hemagglutinin antigens) substances, as well as their concentration and collection; and a conjugation platform providing activation of the virus at a pH that increases binding rate and binding propensity between the virus and the protein, wherein embodiments related to the conjugation platform include controlling the ratio of virus to protein.

Disclosed herein are methods and exemplary compositions associated with antigen purification, exemplary aspects of which may include harvesting viral and antigenic substances from source organisms; and a purification platform comprising chemical separation and size-difference separation for the removal of contaminants, debris and impurities from the viral and protein (e.g. antigenic, including influenza hemagglutinin antigens) substances, as well as their concentration and collection.

Disclosed herein are methods and exemplary compositions associated with virus purification, exemplary aspects of which may include harvesting viral and antigenic substances from source organisms; and a purification platform comprising chemical separation and size-difference separation for the removal of contaminants, debris and impurities from the viral and protein (e.g. antigenic, including influenza hemagglutinin antigens) substances, as well as their concentration and collection.