Temperature-based transient delivery of nucleic acids and proteins to cells and tissues

A method for stimulating an immune response against an antigen in a mammalian subject, comprising:

• administering an effective amount of a composition to the mammalian subject by intradermal injection to stimulate an immune response against the antigen in the mammalian subject,
• wherein the composition comprises an excipient and a temperature-sensitive agent (ts-agent) encoding the antigen, the ts-agent is a temperature-sensitive self-replicating RNA comprising a viral replicon lacking a viral structural protein coding region, the RNA is not packaged in a viral particle, and the ts-agent is capable of expressing the antigen at skin temperature of the subject but not at core temperature of the subject.

A method for stimulating an immune response against an antigen in a mammalian subject, comprising:

• administering an effective amount of a composition to the mammalian subject by intradermal injection to stimulate an immune response against the antigen in the mammalian subject,
• wherein the composition comprises an excipient and a temperature-sensitive agent (ts-agent) encoding the antigen, the ts-agent is a temperature-sensitive self-replicating RNA comprising an Alphavirus replicon lacking a viral structural protein coding region, the ts-agent comprises a nonstructural protein coding region with an insertion of 12-18 nucleotides resulting in expression of a nonstructural Protein 2 (nsP2) comprising from 4 to 6 additional amino acids between beta sheet 5 and beta sheet 6 of the nsP2.

A method for expressing a protein in a mammalian subject, comprising:

• administering an effective amount of a composition to the mammalian subject by intradermal injection to express the protein in the mammalian subject,
• wherein the composition comprises an excipient and a temperature-sensitive agent (ts-agent) encoding the protein, the ts-agent is a temperature-sensitive self-replicating RNA comprising a viral replicon lacking a viral structural protein coding region, the RNA is not packaged in a viral particle, and the ts-agent is capable of expressing the protein at skin temperature of the subject but not at core temperature of the subject.