TY - JOUR ID - 18498 TI - Dystrophin gene editing by CRISPR/Cas9 system in human skeletal muscle cell line (HSkMC) JO - Iranian Journal of Basic Medical Sciences JA - IJBMS LA - en SN - 2008-3866 AU - Dara, Mahintaj AU - razban, vahid AU - Mazloomrezaei, Mohsen AU - Ranjbar, Maryam AU - Nourigorji, Marjan AU - Dianatpour, Mehdi AD - Department of Molecular Medicine, School of Advanced Medical Science and Technology, Shiraz University of Medical Science, Shiraz, Iran AD - Student Research Committee, Shiraz University of Medical Science, Shiraz, Iran AD - Department of Medical Genetics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran Y1 - 2021 PY - 2021 VL - 24 IS - 8 SP - 1153 EP - 1158 KW - CRISPR/Cas9 KW - DMD KW - Dystrophin KW - Gene editing KW - HSkMC DO - 10.22038/ijbms.2021.54711.12269 N2 - Objective(s): Duchene muscular dystrophy (DMD) is a progressive neuromuscular disease caused by mutations in the DMD gene, resulting in the absence of dystrophin expression leading to membrane fragility and myofibril necrosis in the muscle cells. Because of progressive weakness in the skeletal and cardiac muscles, premature death is inevitable.  There is no curative treatment available for DMD. In recent years, advances in genetic engineering tools have made it possible to manipulate gene sequences and accurately modify disease-causing mutations. CRISPR/Cas9 technology is a promising tool for gene editing because of its ability to induce double-strand breaks in the DNA. Materials and Methods: In this study for the exon-skipping approach, we designed a new pair of guide RNAs (gRNA) to induce large deletion of exons 48 to 53 in the DMD gene in the human skeletal muscle cell line (HSkMC), in order to correct the frame of the gene.Results: Data showed successful editing of DMD gene by deletion of exons 48 to 53 and correction of the reading frame in edited cells. Despite a large deletion in the edited DMD gene, the data of real-time PCR, immune florescent staining demonstrated successful expression of truncated dystrophin in edited cells.Conclusion: This study demonstrated that the removal of exons 48-53 by the CRISPR / Cas9 system did not alter the expression of the DMD gene due to the preservation of the reading frame of the gene. UR - https://ijbms.mums.ac.ir/article_18498.html L1 - https://ijbms.mums.ac.ir/article_18498_d988ada858f045a656efe749ff211929.pdf ER -