The INF2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HeLa cells, designed to disrupt the human INF2 gene. This mixed knockout pool provides a convenient loss-of-function model for studying INF2-mediated processes without the need for clonal isolation, preserving population-level heterogeneity that is valuable for pooled screening and functional genomics studies.
The HeLa cell line is an immortalized human epithelial line originating from a cervical adenocarcinoma and is HPV18-positive. Widely used in cell biology, it offers robust growth, ease of genetic manipulation, and a well-characterized signaling context. This background is optimal for investigating actin dynamics, mitochondrial fission, and the interplay between viral oncoproteins and host cell pathways, making it a fitting system to explore INF2 function.
INF2 is a formin-family actin assembly factor that promotes mitochondrial fission downstream of Rho GTPases (RhoA, CDC42) and in response to calcium or ER stress. It nucleates actin filaments at ER-mitochondria contact sites, creating a scaffold that recruits Drp1, the dynamin-related GTPase that drives mitochondrial constriction. INF2 interacts with Spire1 and myosin II, and indirectly with the mitochondrial fission machinery, integrating cytoskeletal dynamics with organelle division. Disruption of INF2 therefore impairs actin-mediated mitochondrial constriction and perturbs ER-mitochondrial tethering.
In the HeLa epithelial cancer cell context, INF2 knockout cells allow dissection of actin-dependent mitochondrial fission in relation to cell migration, division, and stress responses. Since INF2 mutations cause focal segmental glomerulosclerosis (FSGS) and Charcot-Marie-Tooth neuropathy, this model also facilitates research into the cellular mechanisms of these diseases. Although HeLa cells are not podocytes or neurons, they recapitulate the core INF2 interactome and can be used to evaluate mitochondrial morphology defects and to screen compounds that target the INF2?CDRP1 pathway.
Applications include immunofluorescence and MitoTracker staining to assess mitochondrial network architecture, Western blotting and RT-qPCR to confirm gene disruption and monitor expression of RhoA, Drp1, or myosin II, and co-immunoprecipitation to study INF2-containing complexes. The polyclonal format supports high-throughput screening for modulators of actin polymerization or mitochondrial fission. These cells also serve as a valuable system for mechanistic studies of INF2-related pathologies. For inquiries, please contact Ascent Research.
