Investigation of the usability of polymeric nanoparticle based transfection agent for CRISPR/CAS9-mediated editing of COX-2 gene

Genome editing studies are a promising new technology in the scientific world for the treatment of many diseases. The CRISPR/Cas9 system is one of the most common methods used in genome editing studies today. The fact that CRISPR/Cas9 is a simple, fast, and inexpensive method holds great promise for therapeutic applications. Although the CRISPR/Cas9 technology holds great promise, some difficulties must be overcome to be successfully applied in patients. Safe and efficient delivery of the CRISPR/Cas9 genome editing system to target cells in the human body poses one of the biggest challenges of this system. Within the scope of this thesis, the silencing of the COX-2 gene via the CRISPR/Cas9 system was performed in the HUVEC cell line using a PβAE-based transfection agent. Confirmation of silencing of the COX-2 gene in the HUVEC cell line was analyzed by Western blot. In addition, the effects of silencing the COX-2 gene on cell migration in these cells were investigated. As a result, the transfection efficiency of the PβAE transfection agent synthesized and used for the HUVEC cell line was between 90-95% and the CRISPR efficiency was calculated as 57.12% according to the Western blot result. It has been revealed that the PβAE-based nanoparticles used within the scope of this thesis can be used as a good transinfection agent in the HUVEC cell line for CRISPR/Cas9 mediated genome editing, especially for plasmid transfection.