ABHED

When starting our project, our most important goal was to develop a treatment method that could be a hope for all SMA patients. There are many different mutations in the SMN1 gene among SMA patients as well. That’s why we planned our treatment project on the SMN2 gene. Our main goal was that the SMN2 gene produced a functional and 100% full length SMN protein like SMN1, and we tried to understand the exon skipping mechanism associated with it. As a result of our literature searches and analysis on various databases, we have proven in theory that we can achieve this by using the Crispr-Prime editing method. After transferring our Crispr plasmids to the cells we will take from the patient using a lentiviral vector, we have developed a safe treatment method in which we can follow the genome integration and quality control phase, since we will reproduce these cells in culture and give them to the patient. For lentivirus production, we will use PLVX-curo, 3 lentiviral packaging plasmids and pMDlg / PRRE, Prsv-REV and pMD2.G plasmid systems.In order to achieve suitable neuronal motor cells from the fibroblast cells we will take from the patient, we aim to turn these cells into iPS cells under appropriate laboratory conditions after crispr prime editing on fibroblast cells. In both fibroblast cells and iPS cell samples, we will do the p24 ELISA test for showing the absence of RCL. Then we will perform western blot analysis and Sanger DNA sequence analysis to prove that the transferred plasmids do not precede off-target sequences or an oncogene. Finally, after these confirmations were made, we transferred these stem cells to the spinal canal by viral vector injection to the patient.