Investigation of the Usability of Polymeric Nanoparticle-Based Transfection Agent for CRISPR/CAS9 in HUVEC Cell Line

Abstract

Genome editing technology is a promising, popular technology for diagnosing and treating many diseases. One of the most popular techniques utilized in genome editing research nowadays is the CRISPR/Cas system. The CRISPR/Cas system holds considerable potential for diagnostic and therapeutic applications due to its ease of use, speed, and affordability. The safe and effective delivery of the CRISPR-based genome editing system to the intended human body cells is one of the system's problems. This study investigated the silencing of the COX-2 gene via the CRISPR/Cas system using a P?AE-based transfection agent in the HUVEC cell line. Western blot analysis was performed to confirm COX-2 gene silencing in the HUVEC cell line, and the effects of COX-2 gene silencing on cell migration in these cells were examined. As a result, the transfection efficiency of the P?AE-based transfection agent for the HUVEC cell line was determined as 90-95%, and the CRISPR efficiency was calculated as 57.12% based on the Western blot results. It was revealed that the P?AE-based nanoparticles used in this study can be used as a good transfection agent in CRISPR/Cas-mediated genome editing in the HUVEC cell line, especially in plasmid transfection.