METHOD FOR PRODUCING IN VITRO WILSON'S DISEASE MODEL USING HUMAN PLURIPOTENT STEM CELLS, AND IN VITRO WILSON'S DISEASE MODEL PRODUCED THROUGH SAME

摘要

The present invention relates to a method for generating in vitro cultured hepatocytes using an endoplasmic reticulum stress reliever, the method comprising: step A for preparing a guide RNA having a nucleotide sequence of SEQ ID NO: 1; step B for inserting the guide RNA prepared through step A into a Cas9 nuclease expression vector, and thereby preparing a vector for CRISPR/Cas9 cloned so as to include the guide RNA prepared through step A; step C for preparing ssODNs (Single Stranded Oligodeoxynucleotides) having a nucleotide sequence of SEQ ID NO: 2; step D for injecting the vector for CRISPR/Cas9 prepared through step B and the ssODNs prepared through step C into human pluripotent stem cells; step E for obtaining a plurality of single colonies by culturing the human pluripotent stem cells into which the vector for CRISPR/Cas9 and ssODNs have been injected through step D; and step F for sequencing each of the plurality of single colonies obtained through step E, and selecting a single colony having human pluripotent stem cells which have been made able to develop Wilson's disease by being made to include the Wilson's disease-causing gene ATP7B which has mutated into the R778L mutant form through specific gene mutation.