PADI2-mediated citrullination regulates the differences in angiogenic response

Purpose: We have recently identified Padi2 (peptidyl arginine deiminase type II) as a novel angiogenesis-regulating gene by genome-wide association in common inbred mice strains. Here, we further defined the role of PADI2 in angiogenesis and developed both a knock-out (KO) mouse strain and a padi2 null zebrafish in the kdrl: zsGreen transgenic line.Methods: In order to inactivate Padi2 gene, we used the CRISPR-Cas9 system where we designed specific gRNAs against the conserved PADI catalytic domain specific to Padi2 gene in 129S1/SvImJ strain. To determine whether PADI2 modulates angiogenic response in mice, we used the corneal micropocket neovascularization assay with 20ng of bFGF. To generate a mutant padi2 line in zebrafish, we made small deletions within the catalytic domain of padi2 gene also using CRISPR-Cas9 system in the kdrl: zsGreen transgenic zebrafish. Vascular defects were …