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AssigneesSouth Dakota Board of Regents, Cambridge Enterprise Limited, Academisch Ziekenhuis Leiden, University College Cork
InventorsYing Fang, Eric John Snijder, Andrew E. Firth, John F. Atkins, Emma Elisabeth Treffers, Ali Tas, Yanhua Li
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The invention provides the discovery and characterization of a novel arterivirus protein (nsp2TF), whose expression is dependent on −2 ribosomal frameshifting at a site located in the nsp2 coding region. The coding region for the unique TF domain of nsp2TF overlaps the part of ORF1a that encodes the transmembrane region of nsp2 in arteriviruses, including PRRSV, LDV and SHFV. Mutations affecting the expression of nsp2TF impair PRRSV replication and result in a smaller plaque phenotype. Provided herein are arteriviruses that display reduced translation of nsp2TF and/or altered translation of one or more downstream products, arteriviruses in which nsp2TF function is reduced and/or absent, and vaccines comprising said arteriviruses. Also provided herein are diagnostic methods, methods for identifying compounds that inhibit −2 frameshifting, and gene expression tools for eukaryotic systems utilizing −2 frameshifting.
A porcine reproductive and respiratory syndrome virus (PRRSV) comprising a nucleic acid encoding ORF1a or a functional part thereof, wherein said nucleic acid comprises at least one mutation resulting in reduced translation of a transframe (TF) domain of nsp2TF and/or altered translation of one or more downstream products translated from said nucleic acid, in a cell infected by said PRRSV, when compared to a wild-type of said PRRSV, wherein said mutation interferes with −2 ribosomal frameshifting at a −2 frameshifting site located in the nucleic acid sequence that codes for the nsp2 protein GGU(U/C)U(U/C)U and/or the conserved CCCANCUCC motif.
Dependencies are inferred from the claim text.