The INF2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from human embryonic kidney 293T cells, designed to model loss of function of the INF2 gene (inverted formin 2). This polyclonal knockout pool introduces gene disruption via CRISPR/Cas9, generating a heterogeneous mixture of edited alleles that collectively eliminate functional INF2 protein expression. The product offers a versatile and physiologically relevant system for investigating INF2-mediated processes in a well-characterized epithelial cell background.
HEK293T cells are a widely utilized human embryonic kidney cell line that stably expresses the SV40 large T antigen, enabling high-level episomal replication of plasmids containing the SV40 origin of replication. This feature makes HEK293T an ideal host for protein overproduction, viral packaging, and transient transfection studies. The cells exhibit typical epithelial morphology and robust growth characteristics, providing a reliable platform for genetic manipulation and functional assays. Their ease of culture and genetic tractability facilitate reproducible experiments in cytoskeletal and organelle biology.
INF2 functions as an actin nucleation and polymerization factor that regulates both actin cytoskeleton remodeling and mitochondrial fission. It is activated downstream of RhoA, calcium influx, CDC42, and phosphatidylinositol 4,5-bisphosphate (PIP2). Upon stimulation, INF2 promotes actin filament assembly at the mitochondrial outer membrane, which subsequently recruits the dynamin-related protein DRP1 to execute mitochondrial division. INF2 also interacts with myosin, Spire, and FHOD1 to coordinate cytoskeletal dynamics and maintains Golgi apparatus morphology. Disruption of INF2 therefore impairs RhoA-to-actin signaling, DRP1-mediated fission, and overall cytoskeletal architecture, making this knockout a powerful tool for dissecting these interconnected pathways.
In the HEK293T context, loss of INF2 is expected to alter actin polymerization kinetics, mitochondrial network morphology, and cell migration properties. The polyclonal nature of the knockout population avoids clonal artifacts and enables assessment of gene-disruption effects at the population level. Researchers can use these cells to interrogate INF2-dependent pathways under controlled conditions, such as by stimulating with RhoA activators or calcium-mobilizing agents, and monitor outcomes via actin staining, DRP1 localization, and mitochondrial imaging. The model is particularly valuable for differentiating direct INF2 functions from compensatory mechanisms that may arise in clonal isolates.
Typical research applications include investigations of mitochondrial dynamics, actin cytoskeleton regulation, and molecular mechanisms underlying INF2-linked diseases such as focal segmental glomerulosclerosis (FSGS) and Charcot-Marie-Tooth disease (CMT). Experimental assays compatible with these cells encompass western blotting for INF2 and actin, immunofluorescence analysis of mitochondrial morphology, actin polymerization assays, cell migration/invasion tests, DRP1 subcellular localization studies, and calcium flux measurements. These polyclonal knockout cells provide a flexible platform for probing INF2 biology in epithelial cell models. For further technical details and order information, please contact Ascent Research.
