Mutations in chromodomain helicase DNA binding protein 7 (CHD7) gene located in human chromosome 8 have been linked to “CHARGE” syndrome, a cluster of disease seen in a number children, for which there is no pharmacotherapy. As there is no cellular or animal model system to study the function of CHD7, the goal of this investigation is to mimic CHARGE disease in a dish by the use of CRISPR/Cas9 to edit the CHD7 gene in human fibroblast cells. Accordingly, I have generated lentivirus particles encoding Cas9 and CHD7 guide(g)RNA in all-in-one vector, and transduced fibroblast cells. CHD7 mutation will be confirmed by qRT-PCR and DNA sequencing; microscopy, Western blot analyses, and the function of mutant protein will be analyzed. Thereafter, the fibroblast cells that harbor a specific mutation in CHD7 gene will be used to screen drugs using a chemical compound library. Through this assay, a molecule that acts on mutant CHD7 protein as an agonist or antagonist could be found, thereby one could fantasize a therapeutic approach to alleviate CHARGE syndrome. There are limitations to cell-based assays, therefore, animal experiments will be needed to address the toxicity and efficacy of any chemical that might be discovered
