Quantitative analysis of differential expressed genes and pathways in AB8 cells overexpressing Crb2 S267A variant for 48h (+dox) compared to non-induced control cells (-dox) using next-generation sequencing (NGS)

Purpose: NGS can be used for system-based analysis of cellular pathways. The goal of this study was to identify genes and pathways which are activated in AB8 podocytes due to Crb2 S267A overexpression. Methods: AB8 cells were modified using lentiviral transduction with pIND21_hCrb2+GFP_S267A. After selection with puromycin, cells were induced with doxycycline for 48h to overexpress Crb2 S267A variant. Non-induced cells were used as controls. RNA-seq data from AB8 cells overexpressing Crb2 S267A variant for 48h (+dox) were compared with non-induced control cells (-dox) (n=4). Activated pathways (GO terms) were identified with DAVID bioinformatics tool. Results: RNAseq data revealed activation of ER stress response, chaperon folding genes and pathways. Misfolded Crb2 variant induces cellular ER stress compared to control cells. AB8 cells expressing Crb2+GFP_S267A were compared to non-expressing control cells.