Described herein are compositions and methods for tissue-targeted expression of therapeutic genes, using AAV expression vectors that reduce the risk of toxicity associated with AAV gene therapy in the CNS by de-targeting the vulnerable neurons cells including the DRG cells on gene expression, and de-targeting the liver, a major suspect for over-expression in the periphery.

A method for treating cancer by inducing a double strand DNA break at a chromosomal site in cancer cells, inserting a nucleic acid encoding a suicide gene into the chromosomal site having the double strand DNA break, and expressing the suicide gene in the cancer cells, thereby inducing cell death upon expression of the suicide gene. The chromosomal site includes a protospacer adjacent motif (PAM) absent from a corresponding chromosomal site in normal cells. Also disclosed is a cancer treatment method that includes identifying a single nucleotide change in the genomic sequence of cancer cells from a subject as compared to the genomic sequence of normal cells, the single nucleotide change forming a PAM, inserting a suicide gene specifically into the cancer cell genomic sequence, and activating the suicide gene, thereby leading to cell death. Furthermore, provided is a clustered regularly interspaced short palindromic repeat-associated protein having RNA-guided DNA endonuclease activity.

Described herein are chimeric receptors. Chimeric receptors comprise a cytoplasmic domain; a transmembrane domain; and an extracellular domain. In embodiments, the cytoplasmic domain comprises a cytoplasmic portion of a receptor that when activated polarizes a macrophage. In further embodiments, a wild-type protein comprising the cytoplasmic portion does not comprise the extracellular domain of the chimeric receptor. In embodiments, the binding of a ligand to the extracellular domain of the chimeric receptor activates the intracellular portion of the chimeric receptor. Activation of the intracellular portion of the chimeric receptor may polarize the macrophage into an M1 or M2 macrophage.

Monoclonal antibodies and fragments thereof are capable of binding to a serum form of human TK1. Kits and methods involve the use of such monoclonal antibodies or fragments.

Disclosed are the expression of a recombinant chicken IgY scFv antibody and monoclonal antibody raised against human thymidine kinase 1 (hTK1) in prokaryotic cells and a preparation method and use thereof. The scFv antibody includes an amino acid sequence shown in SEQ ID NO: 1. The anti-hTK1 recombinant scFv antibody and monoclonal antibody have good immunoreactivity, high specificity and sensitivity, and are easy to separate and purify.

Embodiments of the disclosure encompass methods and compositions for treatment of cancer utilizing fibroblasts that have been modified to enhance their ability to deliver one or more anti-cancer agents to an individual in need thereof. In particular embodiments, the fibroblasts have been modified to express one or more chemokine receptors and/or have been exposed to hypoxia to enhance their ability to home to cancer cells. In specific cases the modified fibroblasts are engineered to encompass an oncolytic virus as a tumor inhibitory agent.

Embodiments of the disclosure encompass methods and compositions for treatment of cancer utilizing fibroblasts that have been modified to enhance their ability to deliver one or more anti-cancer agents to an individual in need thereof. In particular embodiments, the fibroblasts have been modified to express one or more chemokine receptors and/or have been exposed to hypoxia to enhance their ability to home to cancer cells. In specific cases the modified fibroblasts are engineered to encompass an oncolytic virus as a tumor inhibitory agent.

Nucleic acid sequences encoding improved Herpes Simplex Virus Thymidine Kinases are provided, including their use in diagnostic and therapeutic applications. The thymidine kinases may be mutated using conservative mutations, non-conservative mutations, or both. Also provided are gene therapeutic systems, including viral and retroviral particles.

The present invention relates to a recombinant adenovirus with increased in-vivo safety, tissue specificity, and anticancer activities, and a use thereof. Specifically, the recombinant adenovirus comprising: a promoter of the liver tissue-specific phosphoenolpyruvate carboxykinase (PEPCK) gene; a trans-splicing ribozyme which is operably linked to the promoter and acts on a cancer-specific gene; a therapeutic gene or a reporter gene which is linked to the 3 exon of the ribozyme; and a serotype 35 fiber knob and a serotype 5 shaft, in which the orf4 gene is deleted from adenovirus E1, E3 and E4 orf1, shows remarkable in-vivo safety, high specificity for a target tissue, and remarkable anticancer effects, and thus can be useful for an anticancer drug or a cancer diagnostic agent as a gene delivery vector.

Nucleic acid sequences encoding improved Herpes Simplex Virus Thymidine Kinases are provided, including their use in diagnostic and therapeutic applications. The thymidine kinases may be mutated using conservative mutations, non-conservative mutations, or both. Also provided are gene therapeutic systems, including viral and retroviral particles.