Compositions and methods are provided for the cell-mediated targeted killing of diseased cells based on the presence of an intracellular antigen, rather than a surface-bound marker. The targeting cells are modified to express a cytotoxic protein that is delivered into a targeted cell, and after delivery is selectively activated by the presence of a cytoplasmic protein of interest. In one embodiment of the invention, the cytotoxic molecule is a Granzyme B (GrB) polypeptide. In the compositions of the invention, GrB is modified to render its cytotoxic enzymatic functions inactive, until the presence of an intracellular antigen unlocks the GrB molecule to enable enzymatic activities.

A recombinant vector of a thermolabile UNG fused protein and an expressing and purifying method are provided. The method comprises cloning a Cod UNG genetic sequence to a pCold-SUMO vector to construct a recombinant vector pCold-SUMO-Cod UNG, transforming to E. coli BL21 (DE3) competent cells, transforming and expressing molecular chaperone plasmids pG-Tf2, and inducing the expression at a low temperature to obtain a soluble SUMO-Cod UNG fused protein.

The present invention provides non-natural molecules which comprise a peptide part able to stop the amyloid aggregation which is fused to a moiety which stimulates the proteasomal degradation pathway in the cell. Non-natural molecules of the invention are useful to treat human and veterinary pathological aggregation disorders.

The present invention relates to methods and compositions for reducing the immunogenicity of chimeric Notch receptors, and specifically to transcription factors useful for controlling gene expression delivered to tissues by such chimeric Notch receptors.

The present application relates, in some aspects, to the development of an assay that uses cell survival and/or cell viability as a phenotypic identifier to positively select for agents that destabilize a protein of interest.

In one aspect, the invention provides a peptide comprising a chaperone-mediated autophagy (CMA)-targeting signal domain; a protein-binding domain that selectively binds to a target cytosolic protein; and a cell membrane penetrating domain (CMPD). In another aspect, the invention provides methods for reducing the intracellular expression level of an endogenous target protein in vitro and in an animal, wherein the method involves administration of the peptide. Methods are also provided for treating a pathological condition in an animal, the methods comprising administering the peptide to the animal. In one embodiment, the pathological condition is a neurodegenerative disease. In another embodiment of the invention, the target cytosolic protein is death associated protein kinase 1 and the CMPD is protein transduction domain of the HIV-1 Tat protein.

Systems and methods are presented that allow for selection of tumor neoepitopes that are then used to generate a recombinant polytope that is optimized for proper trafficking and processing. In preferred methods, the polytope is encoded in a viral expression system that is used as a therapeutic agent.

The present disclosure provides regulatable biocircuit systems. Such systems provide modular and tunable protein expression systems in support of the discovery and development of therapeutic modalities.

Expression vectors and methods of protein purification, which allow for selection of full length protein over truncated forms of the protein being purified, are disclosed. The methods express a target protein as a three domain fusion, represented by formula I: A-[L.sub.1]-B-[L.sub.2]-C, where A is a first purification tag domain, C is the second purification tag domain and B is the target protein domain. A, B and C are preferably covalently linked by linkers, L.sub.1 and L.sub.2 as shown in Formula I, however, L.sub.1 and L.sub.2 may be optional. The purification tags at the N and C termini are different.

Expression vectors including nucleic acid sequences which encode the fusion protein represented by formula I are also disclosed. The vectors are used with host expression systems such as insect, yeast, or mammalian cells to express the target protein, which is subsequently purified as a function of the different affinity tags.

The present invention discloses methods and compositions for modulating the quality of an immune response to a target antigen in a mammal, which response results from the expression of a polynucleotide that encodes at least a portion of the target antigen, wherein the quality is modulated by replacing at least one codon of the polynucleotide with a synonymous codon that has a higher or lower preference of usage by the mammal to confer the immune response than the codon it replaces.