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Dara, Mahintaj, razban, vahid, Mazloomrezaei, Mohsen, Ranjbar, Maryam, Nourigorji, Marjan, Dianatpour, Mehdi. (1400). Dystrophin gene editing by CRISPR/Cas9 system in human skeletal muscle cell line (HSkMC). , 24(8), 1153-1158. doi: 10.22038/ijbms.2021.54711.12269
Mahintaj Dara; vahid razban; Mohsen Mazloomrezaei; Maryam Ranjbar; Marjan Nourigorji; Mehdi Dianatpour. "Dystrophin gene editing by CRISPR/Cas9 system in human skeletal muscle cell line (HSkMC)". , 24, 8, 1400, 1153-1158. doi: 10.22038/ijbms.2021.54711.12269
Dara, Mahintaj, razban, vahid, Mazloomrezaei, Mohsen, Ranjbar, Maryam, Nourigorji, Marjan, Dianatpour, Mehdi. (1400). 'Dystrophin gene editing by CRISPR/Cas9 system in human skeletal muscle cell line (HSkMC)', , 24(8), pp. 1153-1158. doi: 10.22038/ijbms.2021.54711.12269
Dara, Mahintaj, razban, vahid, Mazloomrezaei, Mohsen, Ranjbar, Maryam, Nourigorji, Marjan, Dianatpour, Mehdi. Dystrophin gene editing by CRISPR/Cas9 system in human skeletal muscle cell line (HSkMC). , 1400; 24(8): 1153-1158. doi: 10.22038/ijbms.2021.54711.12269

Dystrophin gene editing by CRISPR/Cas9 system in human skeletal muscle cell line (HSkMC)

Iranian Journal of Basic Medical Sciences
مقاله 17، دوره 24، شماره 8، آبان 2021، صفحه 1153-1158    اصل مقاله (685.34 K)
نوع مقاله: Short Communication
شناسه دیجیتال (DOI): 10.22038/ijbms.2021.54711.12269
نویسندگان
Mahintaj Dara1؛ vahid razban1؛ Mohsen Mazloomrezaei2؛ Maryam Ranjbar3؛ Marjan Nourigorji3؛ Mehdi Dianatpour* 3، 4
1Department of Molecular Medicine, School of Advanced Medical Science and Technology, Shiraz University of Medical Science, Shiraz, Iran
2Student Research Committee, Shiraz University of Medical Science, Shiraz, Iran
3Department of Medical Genetics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
4Stem Cell Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
چکیده
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.
کلیدواژه‌ها
CRISPR/Cas9؛ DMD؛ Dystrophin؛ Gene editing؛ HSkMC
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