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.

The present invention relates to modified RDR1 gene capable of conferring virus resistance to a plant and/or increasing virus resistance in a plant, which modification results in enhanced expression of the RDR1 gene, and wherein the modification is selected from a modification that increases the mRNA level of the RDR1 gene; a modification that increases the level of the RDR1 protein; and/or a modification that increases the activity of the RDR1 protein, as compared to a non-modified wild-type RDR1 gene. The modification may comprise a modification upstream of the coding sequence of the RDR1 gene, such as a modification of a regulatory element, preferably of a cis-acting regulatory element. The modified regulatory element is for example selected from a transcription factor binding site for a transcriptional repressor, the modification of which leads to reduction or absence of transcriptional repression; and/or a transcription factor binding site for a transcriptional activator, the modification of which leads to induction or enhancement of transcription; and/or a microRNA binding site, the modification of which leads to reduction or absence of gene repression; and/or a small RNA sequence, the modification of which leads to reduction or absence of gene repression.

The disclosure provides compositions, methods of treatment, and methods of making and using compositions to deliver a nucleic acid to a subject. Compositions described herein include lipid carriers, optionally including an inorganic particle, capable of admixing with nucleic acids. Compositions optionally comprising a nanoparticle carrier and nucleic acid sequence(s) encoding for (i) a cytokine; and (ii) an innate immune stimulator are provided. Further provided are compositions optionally comprising a nanoparticle carrier; an innate immune stimulator; and a nucleic acid sequence encoding for a cytokine. Methods of using the compositions as a therapeutic vaccine for the treatment of a cancer are also provided.

Disclosed herein is a recombinant vector for expressing a replicase comprising a novel polydeoxyribonucleotide. The thus-expressed replicase can be used in in vitro RNA amplification via an RNA-dependent RNA cycling reaction (RCR). Also disclosed herein is a method for producing an amplified RNA product in an RNA cycling reaction (RCR) via use of the present replicase encoded by the present polydeoxyribonucleotide.

Immunogenic compositions and methods of their use in eliciting immune responses to coronaviruses, such as SARS-CoV-2 are provided.

The present invention embraces compositions comprising at least two RNA replicons (self-amplifying RNA vectors (saRNAs or rRNAs)) that can be replicated by a replicase of a self-replicating virus, e.g., a replicase of alphavirus origin. Of the at least two replicons, at least one of which optionally comprises an open reading frame encoding for the RNA-dependent RNA polymerase or replicase that is able to replicate each of the at least two replicons. Further, each replicon comprises an open reading frame encoding for different antigens of interest, e.g., different antigens derived from the same or from different pathogenic organisms, for example the glycoprotein and nucleoprotein of Ebola virus.

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.

The present invention teaches methods and compositions useful for treating, preventing, or curing pathogen infections of living plants. In particular, the present invention teaches methods of enhancing plant response to pathogen-associated molecular patterns using self-amplifying RNA expressing a dsRNA. The methods and compositions described herein are effective at treating biotrophic pathogens, including Liberibacters.

Disclosed are recombinant vectors comprising one or more polynucleotides encoding at least one modified ribonucleic acid (RNA) dependent RNA polymerase (RdRp), as well as pharmaceutical compositions and methods of their use.

The invention relates to a Pseudomonas sp. strain for use in the production of a recombinant protein characterised in that said strain comprises a nucleotide sequence encoding a phi15 RNA polymerase. The invention further relates to a plasmid, capable of integrating or replicating in Pseudomonas sp., comprising a phi15 promoter sequence operably linked to a nucleotide comprising one or more restriction sites for the insertion of a nucleotide sequence encoding a recombinant protein, or operably linked to a nucleotide sequence encoding a recombinant protein.