Disclosed are chimeric stimulatory receptors (CSRs), cell compositions comprising CSRs, methods of making and methods of using same for the treatment of a disease or disorder in a subject.

Provided is a nucleic acid construct for expressing a gene of interest, the nucleic acid construct comprising, in order from 5′-terminus, each sequence of: (a) a 5′ long terminal repeat (LTR) sequence derived from a retrovirus; (b) a packaging signal sequence (IV) derived from a retrovirus; (c) a sequence or multiple cloning site of the gene of interest; (d) a post-transcriptional regulatory sequence (PRE); (e) an siRNA generating sequence which forms at least one stem-loop structure and in which RNA, which induces RNA interference in mammalian cells, is transcribed; and (f) a 3′ LTR sequence derived form a retrovirus.

The present disclosure relates to the field of biotechnology, in particular to a base editing tool and use thereof. The present disclosure provides a fusion protein comprising a first nCas9 fragment, a chimeric insertion fragment, a second nCas9 fragment and two UGI fragments from N-terminus to C-terminus, wherein the chimeric insertion fragment is selected from APOBEC1 fragment or APOBEC3A fragment. The present disclosure provides a novel base editing tool that is compatible with insertion of various deaminases on the chimeric sites of nCas9. Compared with nCas9 terminal fusion base editor, the base editing tool of the present invention significantly reduce of off-targeting on both DNA and RNA, while maintaining specific targeted base editing efficiency, with higher specificity and favorable industrialization prospects.

The present invention provides isolated T cell receptors (TCRs) that specifically bind to an HLA-displayed cancer testis antigen preferentially expressed antigen in melanoma (FRAME) peptide, as well as therapeutic and diagnostic methods of using those isolated TCRs.

Disclosed are an engineered immune killer cell, and a preparation method therefor and the use thereof. The engineered immune killer cell is prepared by inducing reprogrammed human T cell, retains the marker and function of the human T cell from which the engineered immune killer cell is derived, has the marker and function of an NK cell, and transfects and expresses, in an obtained immune killer lymphocyte, a CAR molecule which recognizes tumor and virus-associated antigens or a TCR molecule which specifically recognizes a tumor.

Methods for preparing T cells or NK cells expressing a chimeric antigen receptor (CAR) is described. The methods entail: isolating a population of T cells, generating induced pluripotent stem cells (iPSCs) from the T cells, introducing a nucleic acid molecule encoding a CAR into the iPSCs to create CAR iPSCs; and differentiating the CAR iPSCs into CAR T cells or CAR NK cells.

The present invention relates to a cell resistant to transplantation immune rejection. The cell expresses a first protein recognizing one or more immune effector cells of a host; preferably the cell has the function of inhibiting or killing the immune effector cells of the host. The present invention also relates to a method for preventing or regulating transplantation immune rejection, and a method for preventing or regulating the attack of NK cells on exogenous cells.

Disclosed is an sgRNA guiding a PD1 gene for cleavage to achieve the efficient integration of exogenous sequences. The method for gene editing a PD1 gene in cells includes the steps of introducing a nuclease and an sgRNA into cells, and gene-editing the PD1 gene. The sgRNA guides the nuclease to cleave the PD1 gene and forms a broken site, at which an exogenous donor repair template can also be introduced, so that CAR-T elements can be directionally inserted at the specific site of the PD1 locus to construct enhanced CD19-CART cells with PD1 knockout in one step.

Provided is a chimeric antigen receptor targeting CLL1 and an application thereof. The chimeric antigen receptor targeting CLL1 comprises an antigen binding domain, a hinge region, a transmembrane domain and a signal transduction domain; the antigen binding domain is an anti-CLL1 antibody. The present application uses an anti-CLL1 antibody as the antigen binding domain to construct a chimeric antigen receptor molecule, the chimeric antigen receptor targeting CLL1 has specific targeting effect on CLL1 positive tumor cells, and immune cells expressing chimeric antigen receptor targeting CLL1 have a significant killing effect in vitro and in vivo, and secrete a large amount of cytokine IFN-γ after co-cultured with CLL1 positive tumor cells, which has a specific clearance effect on CLL1 positive tumor cells.

The present invention provides polynucleotide vectors for high expression of heterologous genes. Some vectors further comprise novel transposons and transposases that further improve expression. Further disclosed are vectors that can be used in a gene transfer system for stably introducing nucleic acids into the DNA of a cell. The gene transfer systems can be used in methods, for example, gene expression, bioprocessing, gene therapy, insertional mutagenesis, or gene discovery