Disclosed herein are nucleosides, nucleotides and analogs thereof, pharmaceutical compositions that include one or more of nucleosides, nucleotides and analogs thereof, and methods of synthesizing the same. Also disclosed herein are methods of ameliorating and/or treating a disease and/or a condition, including an infection from a paramyxovirus and/or an orthomyxovirus, with a nucleoside, a nucleotide and an analog thereof.

The present invention provides a recombinant sequence information of a key host factor ANP32A/B which is necessary for the replication of influenza virus in a host. More specifically, the present invention relates to a 129-130 motif and a 149 site of the host factor ANP32A/B protein, which are key active sites for exerting its ability to promote the replication of influenza virus, and are also potential targeting sites of anti-influenza drugs.

Described herein are cell-selective mRNA constructs that can contain a RNA of interest and one or more miRNA targets. The cell-selective mRNA constructs described herein can be used to express an RNA of interest to a cell in a cell-selective manner.

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.

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.

Described herein are cell-selective mRNA constructs that can contain a RNA of interest and one or more miRNA targets. The cell-selective mRNA constructs described herein can be used to express an RNA of interest to a cell in a cell-selective manner.

The present invention enables simultaneous and stable expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8) : (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA expressed in animal cells; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase and of which codons are optimized for the species from which an introduction target cell is derived; (7) an RNA sequence that codes for a protein for regulating the activity of the polymerase and of which codons are optimized for the species from which the introduction target cell is derived; and (8) an RNA sequence that codes for the single-strand RNA binding protein and of which codons are optimized for the species from which the introduction target cell is derived.

The present invention enables simultaneous and stable expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8): (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA expressed in animal cells; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase and of which codons are optimized for the species from which an introduction target cell is derived; (7) an RNA sequence that codes for a protein for regulating the activity of the polymerase and of which codons are optimized for the species from which the introduction target cell is derived; and (8) an RNA sequence that codes for the single-strand RNA binding protein and of which codons are optimized for the species from which the introduction target cell is derived.

Compositions and methods for making cDNA from RNA templates, including double-stranded RNA (dsRNA) and single-stranded RNA (ssRNA), are provided. The composition and methods include a recombinant or purified or modified or recombinant RNA dependent RNA polymerases (RdRp's). The RdRp has an amino acid sequence that shares identity to a contiguous segment of the amino acid sequence of one or more Partitiviridae virus RdRps, and includes a reverse transcriptase (RT) domain. Kits including dNTPs for use in generating cDNAs are included. The RdRps can function efficiently at lower temperatures than previously available RT enzymes. Methods of making cDNAs are provided using an RdRp are also provided. RdRps can also be used to identify candidate reverse transcriptase inhibitors.

Simultaneous expression of a plurality of foreign genes by using a stealthy RNA gene expression system that is a complex that does not activate the innate immune mechanism and is formed from an RNA-dependent RNA polymerase, a single-strand RNA binding protein, and negative-sense single-strand RNAs including the following (1) to (8): (1) a target RNA sequence that codes for any protein or functional RNA; (2) an RNA sequence forming a noncoding region and derived from mRNA; (3) a transcription initiation signal sequence recognized by the RNA-dependent RNA polymerase; (4) a transcription termination signal sequence recognized by the polymerase; (5) an RNA sequence containing a replication origin recognized by the polymerase; (6) an RNA sequence that codes for the polymerase; (7) an RNA sequence that codes for a protein for regulating the activity of the polymerase; and (8) an RNA sequence that codes for the single-strand RNA binding protein.