The present disclosure provides modified cells comprising a nucleic acid encoding an agent (e.g., a detection agent, a selection agent, or a detection agent and a selection agent) inserted at an endogenous proliferation gene or off-target cell marker gene, as well as compositions, methods, uses, and kits related thereto.

Provided herein are systems, compositions, and methods for generating immunogenic peptides or epitopes from tumor associated antigens (e.g., in vivo or ex vivo). Polynucleotides (e.g., genes) encoding the tumor associated antigens may be edited at selected target sites by nucleobase editors comprising a catalytically-inactive Cas9 and a cytosine deaminase, leading to the expression of heteroclitic or cryptic peptides that are more immunogenic than the native peptide derived from the tumor associated antigens. The heteroclitic or cryptic peptide elicit strong tumor-specific immune response (e.g., T-cell response or B-cell response), which inhibits tumor growth and metastasis.

The present invention relates to a recombinant expression vector encoding an inducible cell death switch, a pH-stable fluorophore and a pH-sensitive fluorophore. Moreover, the invention relates to cells comprising said recombinant expression vector as well as their use in an in vitro phagocytosis assay.

The present disclosure is directed to leucine zipper-based sorting systems adapted to facilitate the expression and coordination of polypeptide sequences capable of improving the function of CAR T cells. The systems enable the generation of T cells engineered to express multiple combinations of CARs (multi-CAR), safety-switches, switch receptors, and/or cytokines.

Provided nucleic acid-based expression construct for the target cell-specific production of a therapeutic protein, such as a pro-apoptotic protein, within a target cell, including a target cell that is associated with aging, disease, or other condition, in particular a target cell that is a senescent cell or a cancer cell. Also provided are formulations and systems, including fusogenic lipid nanoparticle (LNP) formulations and systems, for the delivery of nucleic acid-based expression constructs as well as methods for making and using such nucleic acid-based expression constructs, formulations, and systems for reducing, preventing, and/or eliminating the growth and/or survival of a cell, such as a senescent cell and/or a cancer cell, which is associated with aging, disease, or other condition as well as methods for the treatment of aging, disease, or other conditions by the in vivo administration of a formulation, such as a fusogenic LPN formulation, comprising an expression construct for the target cell-specific production of a therapeutic protein, such as a pro-apoptotic protein, in a target cell that is associated with aging, disease, or other condition, in particular a target cell that is a senescent cell or a cancer cell.

The risk with introducing manipulated T-cell is unforeseen adverse events. During the development of chimeric antigen receptor (CAR) T-cell therapies almost all clinical trial has shown some adverse events ranging from cytokine mediated toxicities to tissue damage and death. By the present invention, we aim to induce multiple layers of safety checkpoints. As a last resort we will be able to induce suicide in all cells which we have introduced to the body. Here we describe the scientific background to the parts of our suicide switch and the details regarding the proteins which are included. Safety switches are being tested on CAR-T cells, but they have a few drawbacks. Due to the limited space on the CAR vector, there is only room for one gene switch. Presently the results show they induce apoptosis in around 70-90% of cells, while the desired target is for all cells to be removed from the tissue. By utilising the space available on a synthetic chromosome, we can include multiple genes under Tet operons which allow us to turn multiple genes on/off. Life or death of a cell depends on the balance of the pro and anti-apoptotic proteins. The scale is always shifting a bit back and forth but only when tipped completely over the apoptotic cascade is initiated. By fine tuning the balance we aim to ensure that the hSync carrying cells have a survival advantage in the tumour in the absence of inducing agent. On the other hand, the moment that agent is administrated the cells will undergo apoptosis and express find me and eat me markers making sure they are removed without risk of tissue damage. The present invention encompasses compositions and methods for use in cellular gene therapeutics using a modular approach to genetically engineer cells to carry a synthetic chromosome having a regulatable system including one or more safety switches.

The presently disclosed subject matter provides compositions and systems for cell-based immunotherapy. In certain non-limiting embodiments, the system comprises a membrane-bound polypeptide and at least one soluble polypeptide that is capable of dimerizing with the membrane-bound polypeptide.

An object of the invention is to provide a novel pharmaceutical composition. The pharmaceutical composition of the disclosure contains a DNA encoding a suicide gene having at least one intron sequence. The intron sequence has a donor sequence or an acceptor sequence to be used in a tumor cell with abnormal splicing not in a normal cell. In a transcript of the DNA, the suicide gene is expressed when the intron is abnormally spliced and the suicide gene is not expressed when the intron is not abnormally spliced.

Provided herein are mutants of estrogen receptor alpha ligand binding domain (ER-LBD), and inducible cell death systems that include mutants of estrogen receptor alpha ligand binding domain (ER-LBD). Also provided are methods of for use of the same, such as inducing cell death in a cell.

Provided are an anti-PSMA single-chain antibody, a chimeric antigen receptor associated therewith and use thereof. An amino acid sequence of a heavy chain of the anti-PSMA single-chain antibody includes a sequence shown in SEQ ID NO: 1, and an amino acid sequence of a light chain of the anti-PSMA single-chain antibody includes a sequence shown in SEQ ID NO: 2. The anti-PSMA single-chain antibody is a humanized scFv antibody of PSMA, is more functional in the human body, has better compatibility, and is less prone to be rejected by the immune system. The chimeric antigen receptor has better response effects after specifically bonding to PSMA so that CAR-T cells generate a stronger immune response to tumors, and the chimeric antigen receptor also has better long-term effectiveness than other PSMA chimeric antigen receptors.