The invention provides for immunogenic compositions against West Nile Virus. The immunogenic compositions, in alternate embodiments, also include other equine pathogens. The West Nile Virus composition of the present invention advantageously provides for protection against North American Dominant West Nile Virus strains or isolates.

Certain embodiments are directed to recombinant vesiculovirus encoding a heterologous polynucleotide and methods of using the same.

The present disclosure relates to methods for transiently activating temperature-sensitive agents in one or more cells, for example by contacting one or more cells with a temperature-sensitive agent and transiently incubating the cells at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Additionally, the present disclosure relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent and then lowering the subject's core body temperature to a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. The disclosure also relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent, maintaining the subject's surface body temperature at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Further disclosed are methods of treating a subject with a temperature-sensitive therapeutic agent.

Alkynyl-derivatized cap analogs, alkynyl-modified capped RNA, 1,4-disubstituted triazole-derivatized capped RNA, methods of preparation, methods of isolation, and uses thereof are provided. The click modification facilitates detection and isolation of capped RNAs and the 1,4-disubstituted triazole derivatives formed by the click reaction are useful for producing RNA transcripts and encoded protein.

The present invention discloses an attenuated recombinant alphavirus that is incapable of replicating in mosquito cells and of transmission by mosquito vectors. These attenuated alphavirus may include but is not limited to Western Equine Encephalitis virus, Eastern equine encephalitis virus, Venezuelan equine encephalitis virus or Chikungunya virus. The present invention also discloses the method of generating such alphaviruses and their use as immunogenic compositions.

Certain embodiments are directed to recombinant vesiculovirus encoding a heterologous polynucleotide and methods of using the same.

The present invention discloses an attenuated recombinant alphavirus that is incapable of replicating in mosquito cells and of transmission by mosquito vectors. These attenuated alphavirus may include but is not limited to Western Equine Encephalitis virus, Eastern equine encephalitis virus, Venezuelan equine encephalitis virus or Chikungunya virus. The present invention also discloses the method of generating such alphaviruses and their use as immunogenic compositions.

The invention provides for immunogenic compositions against West Nile Virus. The immunogenic compositions, in alternate embodiments, also include other equine pathogens. The West Nile Virus composition of the present invention advantageously provides for protection against North American Dominant West Nile Virus strains or isolates.

The disclosure provides methods for an in vitro evolution technique to identify and characterize mutations in the non-structural genes of an alphavirus replicon that increase the strength and persistence of expression of the replicon genome. Also provided herein are in vivo methods for administering to an animal model a mutated alphavirus replicon that codes for a gene of experimental or therapeutic interest in the subgenome of the alphavirus replicon. The mutations identified herein improve the therapeutic potential of self-replicating RNA, which may have implications for cancer immunotherapy and beyond, e.g., for vaccination or gene therapy.

The present invention relates to a self-amplifying mRNA vaccine, including: a self-amplifying backbone comprising a live attenuated virus having a hairpin loop insert, and wherein at least one or more structural genes of the live attenuated virus have been replaced with at least one target antigen-encoding nucleic acid sequence.