METHODS AND COMPOSITIONS FOR DIRECTED GENOME MODIFICATION Russian patent published in 2021 - IPC C12N15/85 C12N9/22 C12N5/10 A61K48/00 

FIELD: biotechnology.

SUBSTANCE: present invention relates to biotechnology, in particular to an in vitro method for modifying the genome at a genomic locus of interest in a human induced pluripotent stem cell. To implement the method, a Cas9 protein, an mRNA encoding a Cas9 protein, or a DNA encoding a Cas9 protein, a guide RNA containing CRISPR RNA that hybridizes with the target CRISPR sequence at the genomic locus of interest, and a tracrRNA, or a DNA encoding a guide RNA, are introduced into the specified cell. A large directional vector (LTVEC) is also introduced. In this case, the guide RNA is designed to avoid recognition of any sequence in the nucleotide insert, and after this introduction, the genome of the human induced pluripotent stem cell is modified so that it contains a directed genetic modification.

EFFECT: present invention makes it possible to expand the arsenal of tools for performing precise directed modifications of the genome to identify therapeutic agents.

63 cl, 55 dwg, 48 tbl, 6 ex