Akademik Veri Yönetim Sistemi
Federation of European Biochemical Societies, İstanbul, Türkiye, 5 - 09 Temmuz 2025, ss.457, (Özet Bildiri)
Spinal muscular atrophy (SMA) is the second most common neurodegenerative disease caused by the absence or insufficient of the SMN (survival motor neuron) protein. There are two genes responsible for the SMN protein: telomeric SMN1, which produces 100% functional protein, and centromeric SMN2, which produces only 10% functional protein. Deletion or mutation on SMN1 gene results in insufficient and abnormal protein production and the disease manifests itself. SMN2 copy number is known to have an impact on the clinical severity of the disease. In this study, we examined the effect of growth hormone (GH) on SMN2 promoter activity using reporter in cell culture. Three different sizes of DNA fragments from the promoter region of the SMN2 gene were used to show how gene expression of the reporter construct responds to GH in this study. The first region is 588 bp, second region is 1.036 bp and the third region is 1.705 bp from transcription site of SMN2 gene, respectively. These fragments were amplified by PCR with specific SMN2 primers. SMN2 promoter PCR products were inserted into the pGL3 reporter vector with T4 DNA ligase and the ligation products were transformed into ampicillin plates. Positive colonies containing inserts were identified by direct colony PCR using vector primers. SMN2 promoter inserts in the plasmids were confirmed by PCR with SMN2-specific primers and the plasmids were transfected into CHO cells. After transfection, CHO cells expressing SMN2 promoter driving luciferase gene were treated with growth hormone for 24 hours and then luciferase activity of CHO cells was measured. All reporter constructs containing SMN2 promoters responded to GH. GH induced the highest level of gene expression for the reporter construct containing the 1.036 bp SMN2 promoter. However, GH suppressed gene expression of 1.705 bp SMN2 promoter vector when compared to the gene expression of reporter plasmid alone with GH. GH could be a candidate for the treatment of SMA disease.