Described herein is a method of preventing or treating a disease in a mammalian subject, comprising administering to the subject who is in need thereof an effective dosage of a pharmaceutical composition comprising a virus like particle (VLP) comprising: an alphavirus replicon comprising a recombinant polynucleotide, wherein the polynucleotide comprises a sequence encoding both subunits of a human class II major histocompatibility antigen, a retroviral gag protein, and a fusogenic envelope protein, wherein the VLP does not contain an alphavirus structural protein gene.

The present invention relates to a retrovirus packaging cell which expresses a temperature sensitive RNA-dependent-RNA polymerase (RdRp).

The present invention relates generally to a gene expression system utilizing an alphavirus replicon and T7 promoter. The system is capable of expressing proteins in the cell cytoplasm without integrating the gene of interest into the genome of a host cell. The invention has a wide range of applications such as producing induced pluripotent cells and vaccines against pathogens and cancers.

The invention relates to purified vaccine preparations and methods for providing them. Provided is a method for providing purified viral particles of SFV. comprising the steps of i) providing a preparation of SFV replicon particles: ii) subjecting said preparation to an endonuclease treatment under conditions allowing for degradation of exogenous/host cell DNA and RNA: iii) bringing said endonuclease-treated preparation with a zwitterionic buffer solution to a conductivity of up to about 5.5 mS/cm: iv) contacting the preparation obtained in step (iii) with a strong anion exchange resin: v) eluting the bound SFV replicon particles from said anion exchange resin: vi) bringing the eluted SFV particles to a conductivity in the range of 7.0 to 9.0 mS/cm: vii) contacting the preparation obtained in step (vi) with a strong cation exchange resin under conditions and for a time sufficient to bind to said resin: viii) eluting the bound SFV replicon particles from said cation exchange resin with a zwitterionic buffer solution and collecting at least one fraction containing purified SFV replicon particles: and ix) stabilizing the at least one purified fraction by adding human serum albumin (HSA) to a final concentration in the range of about 0.5-2 w/v %. preferably about 1 w/v %.

Provided is an alphavirus replicon particle (ARP), which comprises (i) alphavirus structural proteins comprising capsid and/or envelope, and (ii) an alphavirus replicon comprising a polynucleotide encoding alphavirus non-structural proteins nsp1, nsp2, nsp3 and nap4 and at least one gene of interest wherein at least one of capsid, and E3 and E2 in the envelope comprise one or more amino acid alteration but E1 in the envelope comprises no amino acid alteration.

The present invention provides nucleic acids sequences, viral particles, viruses, vectors systems, host cells, kits, apparatus, and methods of evolution of a gene product of a gene of interest. The nucleic acid sequences of the invention have been developed primarily for use in evolution of biomolecules of interest. Using a split, non-competent viral vector, the gene of interest can be stably and recombinantly integrated into the viral vector via in-frame insertion of the open reading frame of the gene of interest with an aspect of the native but attenuated viral genome. This configuration of the viral genome, leveraging a split viral vector and an aspect of a viral factor that has been shown to robustly interact with the split viral vector, enables the serial passaging of the recombinant viral particle in an indel and recombination-averse manner.

Provided is an alphavirus replicon particle (ARP), which comprises (i) alphavirus structural proteins comprising capsid and/or envelope, and (ii) an alphavirus replicon comprising a polynucleotide encoding alphavirus non-structural proteins nspl, nsp2, nsp3 and nsp4 and at least one gene of interest wherein at least one of capsid, and E3 and E2 in the envelope comprise one or more amino acid alteration but El in the envelope comprises no amino acid alteration.

The present disclosure relates to the discovery of compositions and methods for therapeutic immunization for SARS-CoV-2 infections and/or disease(s) associated with expression of Glypican-3 (GPC3), including but not limited to cancers such as hepatocellular carcinoma (HCC). Methods of the disclosure include a method of generating virus like vesicles (VLVs), VLVs comprising SARS-CoV-2 antigens from a high titer VLV producing vector, VLVs comprising GPC3 antigens from a high titer VLV producing vector, methods of treating, ameliorating, and/or preventing SARS-COV-2 infection, methods of inducing a memory T and B cell immune response against SARS-CoV-2 infection in a, methods of treating, ameliorating, and/or preventing GPC3 associated disease, and methods of inducing a memory T and B cell immune response against GPC3 in a subject. Furthermore, the disclosure encompasses a pharmaceutical composition for vaccinating a subject to protect the subject against infection with