Abstract: The present disclosure provides a technique whereby the influence of an endogenous TCR is eliminated in TCR gene transfer. A TCR gene is edited using a genome editing enzyme, said genome editing enzyme having one characteristic that amino acids at two specific positions in DNA-binding modules contained in a DNA-binding domain thereof show repeating patterns which differ from one module to another among the four DNA-binding modules. Thus, a lowering in the expression efficiency of the transferred TCR caused by mispairing with an endogenous TCR and the occurrence of a self-reactive TCR are avoided.

Abstract: The present disclosure provides a technique for producing regulatory T cells specific to a desired antigen. Regulatory T cells specific to an antigen are produced by a method that includes: a step for identifying a T cell receptor (TCR) clone present in an effector T cell population specific to an antigen in an effector T cell donor; and a step for introducing all or part of the nucleic acid sequence of the TCR? gene and all or part of the nucleic acid sequence of the TCR? gene included in the clone into a regulatory T cell, said step comprising introducing the TCR? and the TCR? so as to be expressed in pairs.

Abstract: The present disclosure provides a technique for introducing an antigen receptor-encoding nucleic acid into a definite site in a nucleic acid that is contained in an immune cell of a target organism. In the technique according to the present disclosure, a vector, which contains an antigen receptor-encoding nucleic acid, and a nuclease are introduced into an immune cell. The nuclease cleaves the vector and the genome sequence of the immune cell. The vector can be configured so that, when cleaved with the nuclease, a sequence, which is an antigen receptor-encoding sequence and which is introduced by microhomology-mediated end-joining (MMEJ) at the nuclease cleavage site of the genome sequence of the immune cell, is formed.

Abstract: The present disclosure provides a technique for producing regulatory T cells specific to a desired antigen. Regulatory T cells specific to an antigen are produced by a method that includes: a step for identifying a T cell receptor (TCR) clone present in an effector T cell population specific to an antigen in an effector T cell donor; and a step for introducing all or part of the nucleic acid sequence of the TCR? gene and all or part of the nucleic acid sequence of the TCR? gene included in the clone into a regulatory T cell, said step comprising introducing the TCR? and the TCR? so as to be expressed in pairs.

Abstract: The present disclosure provides a technique whereby the influence of an endogenous TCR is eliminated in TCR gene transfer. A TCR gene is edited using a genome editing enzyme, said genome editing enzyme having one characteristic that amino acids at two specific positions in DNA-binding modules contained in a DNA-binding domain thereof show repeating patterns which differ from one module to another among the four DNA-binding modules. Thus, a lowering in the expression efficiency of the transferred TCR caused by mispairing with an endogenous TCR and the occurrence of a self-reactive TCR are avoided.