The ACTA1 Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the DLD-1 human colorectal adenocarcinoma cell line. This product features targeted disruption of the ACTA1 gene, which encodes skeletal muscle alpha-actin, a pivotal cytoskeletal protein. The polyclonal pool offers a heterogeneous loss-of-function model, free from clonal selection artifacts, enabling robust analysis of actin cytoskeleton perturbation in a colorectal cancer background.
DLD-1 cells, established from a colorectal adenocarcinoma, exhibit epithelial morphology and are widely employed as an in vitro model for colorectal cancer research. They harbor mutations in tumor suppressor and oncogene pathways (e.g., APC, KRAS) and display hallmark features of malignancy, including rapid proliferation, adhesion, and migration. This genetic and phenotypic landscape makes DLD-1 an ideal host for interrogating gene function in cancer progression and metastasis.
ACTA1 encodes alpha-skeletal muscle actin, which polymerizes into filamentous actin essential for cell shape, motility, and adhesion. ACTA1 transcription is regulated by SRF/MRTF in response to Rho GTPase and TGF-?? signals. The actin filaments are dynamically remodeled by cofilin, profilin, and the Arp2/3 complex. Alpha-actin directly interacts with myosin II to generate contractile force and associates with vinculin, alpha-actinin, and tropomyosin at focal adhesions, linking to integrin-FAK signaling. Through the RhoA-ROCK-LIMK-cofilin axis, ACTA1 integrates mechanical and chemical cues to control cytoskeletal organization.
In DLD-1 colorectal cancer cells, ACTA1 disruption dismantles actin networks, impairing cell polarity, migration, and invasion. The loss of myosin II contractility and focal adhesion assembly hinders metastastic behaviors, making this knockout model valuable for dissecting actin-dependent mechanisms in epithelial-to-mesenchymal transition and metastatic dissemination. Additionally, the polyclonal composition mirrors tumor heterogeneity, enhancing translational relevance for drug response studies.
These polyclonal knockout cells support a wide range of applications, including western blotting and RT-qPCR for knockout validation, immunofluorescence for F-actin visualization, and transwell migration assays. Co-immunoprecipitation of actin-binding partners and live-cell imaging elucidate dynamic cytoskeletal changes. Drug sensitivity tests can identify actin-targeted therapeutics. This model is also suitable for investigating muscle-specific actin functions in non-muscle neoplasms. For further details, please contact Ascent Research.
