This product consists of CRISPR/Cas9-edited polyclonal A-549 cells carrying a targeted disruption of the ACTN1 gene. The polyclonal knockout population provides a mixed-genotype loss-of-function model, enabling studies of ACTN1 deficiency without clonal isolation. The engineered cells are suitable for investigating the role of alpha-actinin-1 in cytoskeletal organization, focal adhesion dynamics, and cell migration.
The host A-549 cell line is a human lung adenocarcinoma epithelial model established from a 58-year-old Caucasian male. These adherent cells are widely employed in non-small cell lung cancer (NSCLC) research, particularly for studies on tumor progression, metastasis, and chemotherapeutic drug response. A-549 cells harbor a KRAS mutation (G12S) and exhibit an epithelial morphology, making them a relevant system for dissecting signaling pathways that drive oncogenic phenotypes such as migration and invasion.
ACTN1 encodes alpha-actinin-1, an actin-bundling protein that crosslinks filamentous actin and tethers the cytoskeleton to integrin-based focal adhesions. Alpha-actinin-1 directly interacts with vinculin, zyxin, VASP, and integrin beta1, and is regulated by upstream signals from integrin activation, RhoA, Rac1, EGF, and TGF-beta. Downstream, ACTN1 facilitates vinculin and talin recruitment to adhesion sites, promoting FAK and Src phosphorylation, actin polymerization, and cell motility. This positions ACTN1 as a central node in integrin signaling and focal adhesion assembly.
In A-549 cells, disruption of ACTN1 is expected to impair focal adhesion maturation and reduce cytoskeletal tension, leading to attenuated invasive and migratory capacity. This model is therefore highly relevant for studying the mechanisms of NSCLC metastasis, where ACTN1-mediated adhesion and migration are often dysregulated. The knockout cells can be used to dissect the ACTN1-dependent regulation of PI3K-Akt and Rho GTPase pathways, and to evaluate how loss of ACTN1 influences response to targeted therapies or chemotherapeutics in a lung cancer context.
Typical research applications include transwell migration and invasion assays to quantify metastatic potential, immunofluorescence imaging of focal adhesion components (vinculin, paxillin), and phospho-signaling analysis (FAK, Src) to assess pathway activation. Additionally, these cells enable cell adhesion assays and co-culture studies to explore integrin-mediated interactions. The polyclonal format allows for robust, population-level phenotypic analyses without clonal bias. For further details or custom applications, please contact Ascent Research.
