METHOD OF CREATING NEW MUTATIONS IN ORGANISMS AND APPLICATION THEREOF Russian patent published in 2025 - IPC C12N15/10 C12N15/63 C07K14/415 C12N15/82 C12N9/10 C12N9/22 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, namely to a method for creating a new mutation in an organism, which involves the following stages: sequential creation of two or more DNA breaks in a specific site of the organism genome and their spontaneous repair, respectively, where the later DNA break is created based on a new sequence created as a result of the previous repair of the DNA break, to a method of lysis of a DNA target, which includes contact of the DNA target with a complex comprising a Cas polypeptide and at least two targeting RNA, as well as to a method of lysis of a target DNA compositions and a kit for their implementation. Also disclosed is a method of screening editing events independent of an exogenous transgenic marker, involving sequential creation of two or more DNA breaks in a specific site of a first target gene of a recipient cell and spontaneous repair, respectively, where a later DNA break is created on the basis of a new sequence obtained as a result of repair of a previous DNA break, as well as a method for non-transgenic temporary editing of an organism genome.

EFFECT: invention is effective for creating a new mutation in an organism.

26 cl, 29 dwg, 7 tbl, 13 ex