METHODS FOR PRODUCING IN VITRO WILSON'S DISEASE MODEL USING HUMAN PLURIPOTENT STEM CELL AND IN VITRO WILSON'S DISEASE MODEL PRODUCED BY THEREOF

摘要

The present invention relates to a method for generating in vitro cultured hepatocytes using an endoplasmic reticulum stress reliever, comprising: step A of preparing a guide RNA having a nucleotide sequence of SEQ ID NO: 1; Step B of inserting the guide RNA prepared through step A into a Cas9 Nuclease expression-capable vector to prepare a vector for CRISPR/Cas9 cloned to contain the guide RNA prepared through step A; Step C of preparing ssODNs (Single Stranded Oligodeoxynucleotides) having the nucleotide sequence of SEQ ID NO: 2; Step D of injecting the vector for CRISPR/Cas9 prepared through step B and the ssODNs prepared through step C into human pluripotent stem cells; E step of culturing human pluripotent stem cells injected with the CRISPR/Cas9 vector and ssODNs through the D step to obtain a plurality of single colonies; And by performing sequencing for each of a plurality of single colonies obtained through the E step, the Wilson disease causative gene ATP7B modified to the R778L mutant through specific gene mutations, so that the onset of Wilson's disease is possible. It includes; F step of selecting a single colony with induced human pluripotent stem cells.