METHODS OF PRODUCING CELLS EXPRESSING RECOMBINANT RECEPTOR AND RELATED COMPOSITIONS Russian patent published in 2025 - IPC A61K38/00 C12N15/90 C12N15/113 C07K14/705 C07K16/28 A61K35/17 A61P35/00 A61K39/00 

FIELD: biotechnology.

SUBSTANCE: disclosed are methods of producing genetically engineered T-cells, involving: introduction into T-cells of a CRISPR agent containing a guide RNA (nRNA) having a targeting domain which is complementary to at least one target site within exon 1 of the T-cell receptor alpha constant (TRAC) gene and/or a T-cell receptor beta constant (TRBC) gene, wherein said CRISPR agent is capable of causing genetic destruction of at least one target site, which leads to effective endogenous T-cell receptor (TCR) knockout. and introducing a template polynucleotide into said T cells, wherein said template polynucleotide comprises a [5' homology arm]-[transgene]-[3' homology arm]. Also disclosed is a method of producing genetically engineered T-cells, comprising introducing into T-cells having genetic destruction, at least one target site within exon 1 of the alpha T-cell receptor constant (TRAC) gene, leading to an effective endogenous T-cell receptor (TCR) knockout, a template polynucleotide, wherein said template polynucleotide contains a structure [5' homology arm]-[transgene]-[3' homology arm], where said transgene codes a recombinant TCR containing alpha (TCRα) chain and beta (TCRβ) chain, wherein said transgene comprises a nucleic acid sequence coding TCRα chain, and nucleic acid sequence coding TCRβ circuit and kit for implementation of the presented methods.

EFFECT: invention can be used for engineering of immune cells, in particular T-cells.

69 cl, 48 dwg, 10 tbl, 6 ex