Podocyte injury is a convergent driver of glomerular disease progression, yet the genetic determinants that govern podocyte vulnerability to oxidative stress remain incompletely defined. Here, we performed genome-wide CRISPR-Cas9 knockout screens in differentiated human podocytes exposed to two complementary injury models: Adriamycin (ADR), a clinically relevant inducer of podocyte damage, and hydrogen peroxide (H2O2), a direct oxidative stress challenge. Human podocytes were transduced with a genome-wide sgRNA library, differentiated under non-permissive conditions and subjected to optimized stress conditions that imposed selective pressure while preserving library complexity. Across five independent replicates per condition, sequencing-based sgRNA profiling demonstrated high reproducibility, balanced library representation and clear treatment-specific selection. The ADR screen identified 819 depleted and 456 enriched genes, whereas the H2O2 screen identified 277 depleted and 215 enriched genes. Integrated analysis revealed both model-specific dependencies and 27 shared depleted genes across the two injury conditions, implicating stress adaptation, chromatin regulation, mitochondrial metabolism, inflammatory signaling and cytoskeletal organization as candidate processes required for podocyte survival. These genome-wide screens provide a functional framework for prioritizing genes that regulate podocyte resilience to oxidative injury and offer a resource for mechanistic studies of glomerular disease.
