Determination of activation regions of pancreatic transcription factor Pdx1 gene with CRISPR/dCas9

Diabetes mellitus (DM) is a metabolic disorder caused by the absence of insulin hormone or by the inability to influence this hormone. It is known that many genes play a role in the formation of pancreas and differentiation of beta cells. These cells have the potential to differentiate into other types of cells, such as nerve cells, nerve cells, and blood cells. Many methods are used to regulate this gene. Among these, the most common method in recent times is the CRISPR/Cas9 system used by bacteria in the immune response. This system ensures that the desired arrangements are made in the target gene. In the CRISPR/Cas9 system, the CRISPR/dCas9 system, obtained by mutation in the Cas9 protein, allows the regulation of genes. This system allows the expression of the gene to increase (activation) or decrease (suppression) without mutating the target gene. In order for this system to work, however, it must be connected to the dCas9 protein to an activator domain (to increase gene expression) or to a suppressor domain. CRISPR/dCas9-VPR system is the system used in the activation of the desired gene in mammals. Our purpose in this study; Using the CRISPR/dCas9-VPR system, it was to increase the activation of the Pdx1 gene. As a result, significant areas of activation of the Pdx1 gene in the proximal promoter region were determined using CRISPR/dCas9-VPR system.