The INF2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of human HAP1 cells in which the INF2 gene has been disrupted to create a loss-of-function model. This knockout pool enables investigation of inverted formin 2, a key mediator of actin polymerization and mitochondrial fission, without requiring clonal selection. The polyclonal format offers genetic diversity while maintaining functional gene disruption, suitable for robust population-level analyses.
HAP1 is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia cells, displaying an adherent, fibroblast-like morphology. Its haploid genome simplifies CRISPR/Cas9-mediated gene disruption, as most genes exist in a single copy, facilitating efficient knockout generation. Widely employed in functional genomics, HAP1 cells provide a stable and reproducible platform for studying signaling pathways and cellular processes by imaging, biochemical, and pharmacological approaches.
INF2 encodes inverted formin 2, which nucleates actin filaments at endoplasmic reticulum (ER) contact sites. Activated by RhoA GTPase and calcium/calmodulin, INF2 drives actin polymerization that recruits DRP1 and MFF to constrict mitochondria, promoting fission. It interacts with profilin and CDC42 to regulate cytoskeletal dynamics. This pathway lies downstream of RhoA/ROCK signaling, connecting cellular architecture to organelle division. Loss-of-function mutations in INF2 cause focal segmental glomerulosclerosis and Charcot-Marie-Tooth disease, underscoring its role in podocyte and neuronal maintenance.
In HAP1 cells, INF2 knockout impairs formin-mediated actin assembly and disrupts mitochondrial fission, providing a tractable model to dissect RhoA?CINF2?CDRP1 signaling. The polyclonal knockout population allows consistent loss-of-function analysis in a uniform genetic background, ideal for studying cytoskeletal reorganization and mitochondrial network morphology. This system captures key aspects of INF2-dependent cellular pathology relevant to kidney disease, offering a simple yet physiologically meaningful context for mechanistic investigations.
Applications include western blotting and RT-qPCR to validate INF2 disruption, immunofluorescence for actin and mitochondrial markers, MitoTracker staining to assess mitochondrial fragmentation, and actin polymerization assays to measure formin activity. Co-immunoprecipitation can probe interactions with RhoA, profilin, and DRP1. The model is also suitable for drug screens targeting the RhoA?CINF2 axis. For more information or to inquire about customized products, contact Ascent Research.
