IN VITRO EVALUATION CELL MODEL FOR COMPOUND-INDUCED SKIN SENSITIZATION AND CONSTRUCTION METHOD THEREFOR

摘要

An in vitro evaluation cell model for compound-induced skin sensitization and a construction method therefor. The construction method for the cell model comprises the following steps: designing and constructing an sgRNA expression vector by using CRISPR/Cas9; designing and constructing a homologous recombination vector capable of knocking a reporter gene linked with a self-cleavage peptide sequence into the expression frame of HMOX1 gene; and co-transfecting cells with the homologous recombination vector, an hCas9 plasmid and the sgRNA expression vector, and performing monoclonal expansion culture so as to obtain the cell model. An HaCaT cell model in which a luciferase gene has been knocked-in before the stop codon of the HMOX1 gene is obtained by using CRISPR/Cas9 in combination with the cell monoclonal technology . The cell model achieves synchronous expression of the luciferase gene and the HMOX1 gene, thereby effectively distinguishing sensitizing compounds and non-sensitizing compounds.