The ACTA1 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human renal cell adenocarcinoma line, in which the ACTA1 gene encoding alpha-skeletal muscle actin has been disrupted. This heterogeneous pool of knockout cells provides a powerful loss-of-function model for investigating the roles of the actin cytoskeleton in cancer cell biology, without the need for clonal isolation. The polyclonal format ensures representative, bulk-level functional analyses that capture the diversity of gene-edited cells, suitable for a wide range of downstream applications.
The parental 786-O cell line is a widely utilized epithelial model of clear cell renal cell carcinoma (ccRCC), harboring a biallelic VHL tumor suppressor mutation. This deficiency results in constitutive stabilization of hypoxia-inducible factors (HIFs) and drives aggressive tumor phenotypes, including enhanced proliferation, migration, and invasion. As such, 786-O is a standard host for studying the molecular mechanisms of renal cancer metastasis and testing therapeutic strategies.
ACTA1 encodes alpha-skeletal muscle actin, a fundamental component of the actin cytoskeleton that forms filamentous actin (F-actin) networks essential for cell motility, adhesion, and mechanotransduction. In 786-O cells, ACTA1 expression is controlled by the serum response factor (SRF)/myocardin-related transcription factor (MRTF) pathway downstream of RhoA GTPase. ACTA1 interacts with multiple actin-binding proteins, including cofilin, profilin, the Arp2/3 complex, alpha-actinin, and tropomyosin, which together regulate actin polymerization dynamics and higher-order cytoskeletal structures. Disruption of ACTA1 compromises focal adhesion assembly, reducing integrin-mediated signaling through FAK and paxillin, attenuating Rho GTPase (RhoA, Rac1) activity, and impairing mechanosensitive YAP/TAZ nuclear localization.
The combination of ACTA1 knockout with the VHL-mutant 786-O background creates a physiologically relevant model for dissecting the contribution of actin cytoskeletal remodeling to renal cell carcinoma invasion and metastasis. The loss of alpha-skeletal muscle actin destabilizes the actin network, leading to reduced cell adhesion strength and directional migration??key processes hijacked during tumor dissemination. This model uniquely facilitates investigation into cross-talk between hypoxia-driven HIF pathways and actin-mediated signaling nodes, helping to identify actin-dependent vulnerabilities in ccRCC.
These polyclonal knockout cells can be employed in a variety of functional assays. Researchers can perform Western blotting to quantify changes in actin and associated proteins, immunofluorescence staining to visualize F-actin reorganization, wound healing and transwell migration assays to assess cell motility, and cell adhesion assays under different matrix conditions. Additional applications include G-LISA activation assays for RhoA and Rac1, co-immunoprecipitation to map altered interactions with actin-binding partners such as cofilin or Arp2/3, and pharmacological studies with cytoskeletal inhibitors like latrunculin or cytochalasin. For further information or to place an order, please contact Ascent Research.
