The invention provides pluripotent cells that are used therapeutically for regenerating tissues but avoid rejection by subjects that receive them. In particular, the invention provides hypo-immunogenic pluripotent cells that avoid host immune rejection. The cells lack major immune antigens that trigger immune responses and are engineered to avoid phagocytic endocytosis. The invention further provides universally acceptable “off-the-shelf” pluripotent cells and derivatives thereof for generating or regenerating specific tissues and organs.

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 disclosure provides an engineered stem cell, comprising a vector comprising a polynucleotide comprising a nucleic acid sequence of suicide gene, a nucleic acid sequence of immune checkpoint gene and a natural cytotoxicity triggering receptor or a TNF-related apoptosis-inducing ligand, wherein the stem cell is a tumor-targeting cell. The present disclosure also provides a method for treating a cancer or enhancing intratumor immunity or enhancing immunity in tumor microenvironment in a subject, comprising administering an effective amount of the engineered stem cell of the present disclosure to the subject.

Disclosed are methods of treating a cancer in a subject, comprising treating the subject with a combination of gene-mediated cytotoxic immunotherapy and a DNA damage response inhibitor.

Some embodiments of the methods and compositions provided herein relate to chimeric cytokine receptors. In some embodiments, a chimeric cytokine receptor can include an IL-7 tethered to an extracellular IL-7 receptor domain, and an intracellular IL-21 receptor domain linked to the extracellular IL-7 receptor domain. In some embodiments, a T cell containing a chimeric cytokine receptor can be readily activated and/or expanded in the absence of an exogenous cytokine.

Provided are compositions and methods for using the compositions to modify eukaryotic chromosomes. The methods involve iteratively inserting DNA payloads into a chromosomal locus, or into multiple chromosomal loci. The methods utilize positive and negative selection approaches in combination with one or more recombinases to select cells that contain a payload, eliminate cells that do not contain a payload, and sequentially replace contiguous segments of the chromosome with subsequent payload insertions. Modified cells, and modified mammals containing modified cells, are included.

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.

Disclosed are a kit and use thereof. The kit includes a first polyclonal antibody that has been immobilized or suitable for immobilization on a solid carrier, a second polyclonal antibody labeled with a marker. Both the first polyclonal antibody and the second polyclonal antibody are both chicken anti human-thymidine kinase IgY-polyclonal antibodies. Both the first polyclonal antibody and the second polyclonal antibody are suitable for specifically binding to thymidine kinase 1. The kit is suitable for the risk assessment of micro malignant tumors/precancerous diseases and tumors that are not detectable by human population screening images.

The present disclosure provides nucleic acid constructs for the treatment of cancer, comprising a cancer-specific promoter and one or more therapeutic genes.

The invention relates generally to a method of treatment for a human genetic disease, such as diseases characterized by unbalanced nucleotide pools, e.g., mitochondrial DNA depletion syndromes, and more specifically, thymidine kinase 2 (TK2) deficiency, using gene therapy. The gene therapy may involve administration of one or more constructs, such as a viral vector, containing a nucleic acid encoding a functional protein. The functional protein may correspond to a nuclear gene. For treatment of TK2 deficiency, the gene therapy may involve administration of one or more constructs, such as a viral vector, containing a nucleic acid encoding a functional TK2 enzyme. The treatment may also involve the administration of pharmacological therapy in conjunction with the gene therapy. The treatment protocols of the disclosure, such as those involving gene therapy alone or in combination with pharmacological therapy, can be used to treat, prevent, and/or cure various other disorders of unbalanced nucleoside pools, especially those found in mitochondrial DNA depletion syndrome.