Abstract: An adenovirus expression vector is provided. The adenovirus expression vector may include: a) one or more mutations that render the adenovirus replication incompetent and b) at least one nucleotide sequence encoding a protein or an RNA is provided. A method of synthesizing an adenovirus vector is also provided. The synthesis may include: a) producing a plurality of overlapping adenovirus sub-fragments, each sub-fragment comprising a portion of the full genome of the adenovirus; b) circularizing the sub-fragments to form plasmid structures; and c) assembling the circularized sub-fragments into a linear structure, wherein the vector comprises a combination of two or more sub-fragments. A mammalian cell line configured to replicate adenoviral vectors, wherein the cell line comprises nucleotide sequences expressing E1A and E1B gene products but is devoid of other adenovirus sequences is also provided.

Abstract: The present disclosure relates to CCL20 promoters derived from the regulatory region of the CCL20 gene that stimulate gene expression in response to inflammation. These CCL20 promoters, which can be stimulated by an endogenous or exogenous cytokine, can be used to control the timing of the expression of a heterologous gene. The present disclosure also relates to expression cassettes or vectors comprising the CCL20 promoters of the present disclosure operably linked to a nucleic acid sequence encoding, e.g., a polypeptide of interest, as well as delivery systems (e.g., viral particles, lipid vesicles, or nanoparticles) or cells comprising such expression cassettes or vectors. The present disclosure also relates to the use of the promoters, expression cassettes, vectors, delivery systems, or cells in the treatment of diseases related to inflammation, or for recombinant gene expression.

Abstract: Provided are methods, polynucleotides and compositions for the generation of engineered Treg cells. The methods, polynucleotides and compositions enable the reprogramming of hematopoietic cells into Treg cells by constitutive or regulated expression of FOXP3 in engineered cells such that the engineered Treg cells can suppress the activation and proliferation of responder T cells.

Abstract: Modular synthetic receptors are provided. The synthetic receptors may include an extracellular domain capable of binding to one or more ligand molecules and may be released from the synthetic receptor after binding, a transmembrane domain derived from the Notch receptor, and an intracellular domain which may have one or more functional activities when released from the synthetic receptor. A method of use for the synthetic receptors is also provided, wherein upon binding of the extracellular domain to a specific ligand, the synthetic receptor undergoes proteolytic cleavage to release either or both the extracellular and intracellular domains. The extracellular binding domain, if released, may continue to bind to its cognate ligand and may carry one or more additional functional activities and the intracellular domain, if released, may stimulate or inhibit one or more intracellular activities.

Abstract: The present disclosure relates to bicistronic polypeptide constructs for use in allogeneic gene therapy, for example, CAR-T cell therapy. The bicistronic constructs comprise first polynucleotide encoding a therapeutic molecule (e.g., a CAR-T or an antibody) and a second polynucleotide encoding an Immune Surveillance Masking Molecule (ISMM). The ISMM comprises a human leukocyte antigen-E genetically fused to a non-functional version, e.g., a fragment, of the protein knock-out by the insertion of the bicistronic construct, e.g., beta-2 microglobulin or B2M. Also provided are vectors comprising the bicistronic constructs, cells (e.g., CAR-T cells), and methods of use. Also provided are kits and articles of manufacture. The present disclosure also provides four novel insertion sites that can be used to insert an expression construct in the B2M gene.

Abstract: Modular synthetic receptors are provided. The synthetic receptors may include an extracellular domain capable of binding to one or more ligand molecules and may be released from the synthetic receptor after binding, a transmembrane domain derived from the Notch receptor, and an intracellular domain which may have one or more functional activities when released from the synthetic receptor. A method of use for the synthetic receptors is also provided, wherein upon binding of the extracellular domain to a specific ligand, the synthetic receptor undergoes proteolytic cleavage to release either or both the extracellular and intracellular domains. The extracellular binding domain, if released, may continue to bind to its cognate ligand and may carry one or more additional functional activities and the intracellular domain, if released, may stimulate or inhibit one or more intracellular activities.

Abstract: Modular synthetic receptors are provided. The synthetic receptors may include an extracellular domain capable of binding to one or more ligand molecules and may be released from the synthetic receptor after binding, a transmembrane domain derived from the Notch receptor, and an intracellular domain which may have one or more functional activities when released from the synthetic receptor. A method of use for the synthetic receptors is also provided, wherein upon binding of the extracellular domain to a specific ligand, the synthetic receptor undergoes proteolytic cleavage to release either or both the extracellular and intracellular domains. The extracellular binding domain, if released, may continue to bind to its cognate ligand and may carry one or more additional functional activities and the intracellular domain, if released, may stimulate or inhibit one or more intracellular activities.

Abstract: An adenovirus expression vector is provided. The adenovirus expression vector may include: a) one or more mutations that render the adenovirus replication incompetent and b) at least one nucleotide sequence encoding a protein or an RNA is provided. A method of synthesizing an adenovirus vector is also provided. The synthesis may include: a) producing a plurality of overlapping adenovirus sub-fragments, each sub-fragment comprising a portion of the full genome of the adenovirus; b) circularizing the sub-fragments to form plasmid structures; and c) assembling the circularized sub-fragments into a linear structure, wherein the vector comprises a combination of two or more sub-fragments. A mammalian cell line configured to replicate adenoviral vectors, wherein the cell line comprises nucleotide sequences expressing E1A and E1B gene products but is devoid of other adenovirus sequences is also provided.