TALEN and CRISPR/Cas9 editing NF-κB gene to construct stable transgenic cell lines

In order to effectively and rapidly screen NF-κB inhibitors, all five genes of the NF-κB family （RELA、 RELB、 CREL、NF-κB1、and NF-κB2） in three different cell lines （293H, Huh7, and CAPAN-1） were edited using TALEN and CRISPR/Cas9. The edited NF-κB gene was repaired using a linear homologous arm donor containing the EGFP gene sequence , the gene editing efficiency was simply determined by observing the intensity of intracellular fluorescence expression, and the editing efficiency was also detected by fluorescence quantitative PCR of NF-κB-EGFP fusion genes. It was found that the editing efficiency of TALEN is 1.67 times, 1.58 times, and 1.65 times higher than that of CRISPR/Cas9 in 293H, Huh7, and CAPAN-1 cells, respectively. Finally, positive cells were isolated from TALEN edited cells using flow cytometry and cultured and verified, The use of BAY 11-7821 and miR223 significantly reduced the number of fluorescent cells in 293H cells, with average inhibition rates of 76.6% and 66.4%, respectively. This study establishes three stable cell line platforms for highly effective screening NF-κB inhibitors.