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OwnerAnne Elizabeth SIMON, Chanyong JANG +5inventors
InventorsAnne Elizabeth SIMON, Chanyong JANG, Elizabeth CARINO, Feng GAO, George BELOV, Anna ZIMINA, Stephen Xiaohua YANG
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The present disclosure relates to a viral vector having an exogenous RNA segment with a hairpin-like structure, for example having two or more base-paired regions separated by one or more non-base-paired regions. The exogenous RNA segment may have a secondary structure, minimum free energy, average positional entropy or other attributes within specified ranges, or with values similar to one or more hairpin-like structures of a reference wild type virus. In some examples, the viral vector is a live attenuated vaccine. In some examples, the viral vectors downregulates a susceptibility gene in a host plant.
A ribonucleic acid (RNA) molecule derived from an RNA virus whose nucleotide sequence has been modified to comprise a heterologous RNA segment, wherein the heterologous RNA segment comprises two or more base paired regions, one or more non-base-paired regions separating the base paired regions, and an apical loop at the end of one of the base-paired regions, and wherein the heterologous RNA segment has an average position entropy (APE) in the range of 0.01 and 0.75 and a minimum free energy in kcal/mol in a range of about −5 to +15, or in a range of about −10 to +10, from −0.44 multiplied by the number of bases of the heterologous RNA segment minus 1.89 or, if the heterologous RNA segment has more than 150 nucleotides, a minimum free energy in kcal/mol in a range of about −20 to +20 from −0.44 multiplied by the number of bases of the heterologous RNA segment minus 1.89.
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A ULV, ulaRNA, Class 2 ULV, CY1, CTV or TRV vector comprising a heterologous RNA segment targeted against a host plant susceptibility gene, wherein the heterologous RNA segment mimics one or more wildtype hairpin-like regions of the vector and/or wherein the heterologous RNA segment comprises two or more base paired regions, one or more non-base-paired regions separating the base paired regions, and an apical loop at the end of one of the base-paired regions, and wherein the heterologous RNA segment has an average position entropy (APE) in the range of 0.01 and 0.75 and a minimum free energy in kcal/mol in a range of about −5 to +15, or in a range of about −10 to +10, from −0.44 multiplied by the number of bases of the heterologous RNA segment minus 1.89 or, if the heterologous RNA segment has more than 150 nucleotides, a minimum free energy in kcal/mol in a range of about −20 to +20 from −0.44 multiplied by the number of bases of the heterologous RNA segment minus 1.89.
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A method of attenuating a live RNA virus vaccine comprising a step of providing a heterologous RNA insert, wherein the heterologous RNA insert is complementary to a microRNA of a host animal and the heterologous RNA insert comprises two or more base paired regions, one or more non-base-paired regions separating the base paired regions, and an apical loop at the end of one of the base-paired regions, and wherein the heterologous RNA segment has an average position entropy (APE) in the range of 0.01 and 0.75 and a minimum free energy in kcal/mol in a range of about −5 to +15, or in a range of about −10 to +10, from −0.44 multiplied by the number of bases of the heterologous RNA segment minus 1.89 or, if the heterologous RNA segment has more than 150 nucleotides, a minimum free energy in kcal/mol in a range of about −20 to +20 from −0.44 multiplied by the number of bases of the heterologous RNA segment minus 1.89.
Dependencies are inferred from the claim text.