The ACTR3C Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney epithelial cell line, engineered to disrupt the ACTR3C gene. This targeted gene disruption eliminates functional ACTR3C, a core component of the Arp2/3 complex, providing a loss-of-function model to dissect the complex’s roles in actin cytoskeletal organization, cell motility, and intracellular trafficking. As a polyclonal population, these cells offer a genetically heterogeneous knockout background suitable for pooled functional assays and robust phenotypic screening, without confounding monoclonal artifacts.
The HEK293T host cells are a widely adopted model system stably expressing the SV40 large T antigen, which facilitates episomal replication of plasmids containing the SV40 origin of replication and enables high-level recombinant protein expression. This cell line is a mainstay for transient transfection, viral vector production, and protein biochemistry due to its ease of culture and transfection efficiency. The epithelial origin of HEK293T cells renders them particularly relevant for investigating actin-dependent processes such as adhesion, migration, and membrane dynamics in a controlled cellular environment.
ACTR3C is a structural subunit of the heptameric Arp2/3 complex, the primary nucleator of branched actin networks. Upon activation by Rac1, Cdc42, and nucleation-promoting factors (WASP, N-WASP, WAVE complex), the Arp2/3 complex initiates actin filament branching, driving lamellipodia and filopodia protrusion. The complex interacts with actin filaments and is regulated by upstream signals from EGFR and PDGFR via PI3K. Downstream, it influences focal adhesion dynamics, endocytic vesicle formation, and myosin II contractility, with cofilin and profilin modulating actin turnover. ACTR3C disruption thus perturbs Rho GTPase-to-cytoskeleton signaling.
In the HEK293T background, ACTR3C knockout is predicted to compromise actin filament branching and lamellipodia formation, leading to impaired cell migration, attenuated integrin-mediated adhesion turnover, and reduced endocytic capacity. Given the cell line’s utility in viral production, actin defects may also affect viral entry and egress pathways that exploit host cytoskeletal machinery. This polyclonal knockout model therefore allows researchers to interrogate the specific contribution of ACTR3C to Arp2/3 complex assembly and activity in epithelial cells, and to screen for phenotypic modifiers without the confounding influence of basal ACTR3C expression.
Typical applications include Western blotting and co-immunoprecipitation to assess Arp2/3 complex composition, phalloidin staining and immunofluorescence to visualize actin architecture, and functional assays such as wound healing and Transwell migration to measure motility. This model supports studies of cancer metastasis, endocytosis, and cytoskeletal drug screening. Live-cell imaging can reveal actin dynamics. For further details, please contact Ascent Research.
