The present invention relates to a method for producing eukaryotic cell lysates for cell-free protein synthesis comprising at least one exogenous enzyme, wherein the method comprises at least the following steps: a) providing eukaryotic cells transfected with at least one donor template coding for at least one exogenous enzyme which is selected from the group comprising orthogonal aminoacyl-tRNA synthetases and viral RNA polymerases: b) cultivating the transfected cells of step a) for a predetermined period of time and subsequently harvesting the cells; and c) disrupting the harvested cells and preparing a cell lysate for cell-free protein synthesis therefrom. More specifically, the invention relates to a method for producing eukaryotic cell lysates which are capable of cell-free synthesis of a target protein comprising a non-canonical amino acid, wherein the method comprises at least the following steps: a) providing eukaryotic cells transfected with at least one donor template coding for an orthogonal aminoacyl-tRNA synthetase. i.e. an aminoacyl-tRNA synthetase which is specific for a tRNA which is not recognized by endogenous aminoacyl-tRNA synthetase in said eukaryotic cells and is specific for a corresponding non-canonical amino acid: b) cultivating the transfected cells of step a) for a predetermined period of time and subsequently harvesting the cells; and c) disrupting the harvested cells and preparing a cell lysate therefrom. Further aspects of the present invention relate in particular to eukaryotic cell lysates obtainable by the above methods as well as to a method for performing cell-free synthesis of a target protein comprising a non-canonical amino acid.

The invention provides methods of making vaccines against viruses, including against SARS-Cov-2. Such methods entail identifying areas of a viral genome that are highly conserved and making vaccines that target the highly conserved areas. The invention provides a polypeptide vaccine comprising a SARS-Cov-2 polypeptide or an immunogenic fragment thereof and a pharmaceutically acceptable excipient. The invention provides a polynucleotide vaccine comprising a polynucleotide encoding a SARS-Cov-2 polypeptide or immunogenic fragment thereof linked to a heterologous promoter and a pharmaceutically acceptable excipient. The invention provides methods for effecting prophylaxis of or treating SARS-Cov-2 infection comprising a step of administering a polypeptide vaccine and/or a polynucleotide vaccine to a subject in need thereof.

The disclosure provides nanoparticle and compound compositions and methods of making and using the same to a nucleic acid encoding a protein, antibody, or functional fragment thereof for administration to a subject. Various nanoparticle carriers are described. In some instances, the nanoparticle component may include a hydrophobic core having an inorganic particle, and optionally a membrane having a cationic lipid.

The present invention provides a live, attenuated coronavirus comprising a variant replicase gene encoding polyproteins comprising a mutation in one or more of non-structural protein(s) (nsp)-10, nsp-14, nsp-15 or nsp-16. The coronavirus may be used as a vaccine for treating and/or preventing a disease, such as infectious bronchitis, in a subject.

The present application provides an RNA-based system for generating a virus of the Orthomyxoviridae family, and methods of producing viruses of the Orthomyxoviridae family using such system.

The present invention relates to a genetic determinant which may comprise at least two copies of a combination of two closely linked RDR1 genes, which two closely linked RDR1 genes are inversely oriented, and which genetic determinant leads to virus resistance when present in a plant. In one embodiment, of the RDR1 genes in the combination is represented by SEQ ID No. 1 or has at least 70% sequence identity, and one of the RDR1 genes in the combination is represented by SEQ ID No. 3 or has at least 70% sequence identity; or one of the RDR1 genes in the combination encodes a protein represented by SEQ ID No. 2 or a protein that has at least 70% sequence identity, and one of the RDR1 genes encodes a protein represented by SEQ ID No. 4 or a protein that has at least 70% sequence identity.

Provided herein are attenuated influenza viruses and methods of making attenuated influenza viruses.

This document relates to methods and materials involved in treating and/or preventing and/or reducing the severity of a viral infection present in a mammal. For example, methods and materials for reducing the severity of a viral infection present in a mammal (e.g., a human) are provided.

Provided herein are attenuated influenza viruses and methods of making attenuated influenza viruses.

The present disclosure provides compositions and methods for the targeted modification of RNA molecules by RNA prime editing. The compositions and methods may be conducted invitro or in vivo within cells (e.g., human cells) for the therapeutic correction of disease-causing mutations and/or installation of motifs or mutations in RNA molecules of interest as a tool for scientific research. The disclosure provides compositions and methods for conducting RNA prime editing of a target RNA molecule (e.g., an RNA transcript) that enables the incorporation of one or more nucleotide changes and/or targeted mutagenesis of a target RNA molecule. The nucleotide change can include a single-nucleotide change, an insertion of one or more nucleotides, or a deletion of one or more nucleotides. More in particular, the disclosure provides a variety of configurations of the RNA prime editors each comprising a nucleic acid programmable RNA binding proteins (napRNAbp), such as Cas13, and an RNA-dependent RNA polymerase (RDRP), which are provided as fusion proteins or which can be separately provided in trans. The RNA prime editors are guided to a target RNA site by a guide RNA, which can be a rpegRNA that includes a template region for the synthesis of an RNA sequence to be installed on the RNA molecule attached to an available 3 terminus. In others embodiments, the RNA template can be provided in trans.