SARS-CoV-2 proteins inhibit mitochondrial HIGD1A to induce complement-mediated endothelial cell injury

Purpose: The purpose is to systemically identify the genes regulating CD59 expressed on endothelial cells by Crispr/cas9-gRNA library screening Methods: We profiled gRNA sequnecing of endothelial cells with significant CD59 changes and those cells without significant CD59 changes Results: several genes with upregulated or downrgulated were identified as regulators of CD59 in endothelial cells with the edited genome Overall design: Endothelial cell lines (HMEC-1) were transfected with lentivirus packaged with Cripsr/Cas9-gRNA library at a MOI of 0.3; after puromycin selection, surviving cells were cultured into many wells, two wells were transfected with null plasmid (Mock), some wells were transfected with N protein plasmid, and some wells were transfected with a pooled plasmids repectively SARS-CoV-2 proteins;half of wells were cultured regularly and half of wells were cultured in high glucose condition. After 48-72h, the expression of CD59 was detected. Cells had significant CD59 changes were subjected into the second screening as the first one. Cells had no significant CD59 changes were pooled as control group, and cells with significant CD59 changes after two cycles of screening were pooled as experiment group. We then performed gRNA sequencing of the two groups.