METHODS AND COMPOSITIONS FOR PRODUCING OR MAINTAINING PLURIPOTENT CELLS Russian patent published in 2019 - IPC C12N5/74 C12N5/00 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, specifically to producing a population of human induced pluripotent stem cells (hiPSC), maintaining hiPSC, modifying the target genomic locus in hiPSC and a composition for culturing and maintaining hiPSC. Method includes in vitro cultivation of a fibroblast population transformed to express a pluripotent state in a medium with low osmolality of 200 mOsm/kg to 250 mOsm/kg containing a basic medium and a polypeptide which is a leukemia inhibiting factor (LIF), glycogen-synthase-kinase 3 (GSK3) inhibitor and a MEK inhibitor. Modification of the target genomic locus in the hiPSC is performed by introducing into the hiPSC a targeting vector comprising a polynucleotide insert flanked by 5'- and 3'-homologous arms corresponding to 5'- and 3'-initial sites in the target genomic locus or introduction into the hiPSC of the nuclease agent, which creates one or more single-stranded or double-stranded breaks in the recognition site in the target genomic locus to create modification of target genome locus and identification of genetically modified hiPSC containing in its genome polynucleotide insert built into target genomic locus.

EFFECT: invention widens the range of equipment.

38 cl, 6 tbl, 6 dwg, 5 ex