This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HT29 human colorectal adenocarcinoma cell line, targeting the ARPC1A gene. ARPC1A encodes the p41-Arc subunit of the Arp2/3 complex, a critical nucleator of branched actin filaments. The polyclonal knockout population provides a heterogeneous loss-of-function model suitable for studying ARPC1A-dependent cellular processes in a colorectal cancer background. This product is designed for researchers investigating actin cytoskeleton dynamics, cell migration, and related signaling pathways.
HT29 is an epithelial colorectal adenocarcinoma cell line derived from a 44-year-old female. It harbors oncogenic mutations including BRAF V600E, APC, and TP53 mutations, while maintaining wild-type KRAS status. This genetic profile renders HT29 a well-established in vitro model for colorectal cancer research, particularly for studying MAPK pathway-driven tumorigenesis and metastasis. The cell line??s epithelial origin and adherent growth characteristics make it suitable for imaging-based assays of cell morphology and motility.
The ARPC1A protein is an essential component of the heteroheptameric Arp2/3 complex, which nucleates actin polymerization to generate branched F-actin networks. This activity is stimulated by nucleation-promoting factors such as WAVE2 downstream of Rac1 and Cdc42 GTPases. ARPC1A interacts directly with other Arp2/3 subunits (ARPC2, ARPC3, ARPC4, ARPC5, ARP2, and ARP3) and with cortactin. ARPC1A-dependent branched actin assembly is critical for lamellipodium formation, cell migration, endocytosis, and focal adhesion dynamics. Disruption of ARPC1A is expected to impair WAVE-mediated actin reorganization and downstream cellular processes.
In the HT29 colorectal cancer context, ARPC1A knockout disrupts actin-based protrusive structures and cell motility, providing a powerful tool to dissect the role of branched actin nucleation in tumor cell invasion and metastasis. Given the BRAF V600E-driven background, this model is particularly relevant for investigating how oncogenic signaling intersects with actin cytoskeleton regulation. The polyclonal nature of the knockout population mimics heterogeneous gene disruption, enabling the study of population-level effects on migration and invasion without clonal bias. This model can be employed to screen anti-migratory compounds and to evaluate the impact of ARPC1A loss on epithelial-mesenchymal transition (EMT) programs.
Researchers can employ these polyclonal ARPC1A knockout HT29 cells in various assays, including transwell migration and invasion assays, wound-healing scratch assays, and time-lapse live-cell imaging to quantify motility defects. F-actin architecture can be visualized by phalloidin staining, while western blotting and co-immunoprecipitation facilitate analysis of Arp2/3 complex integrity and interactions. The model supports studies of WAVE regulatory complex signaling, Rho GTPase-mediated actin remodeling, and endocytic trafficking. It is also suitable for anti-metastatic drug screening and functional interrogation of cytoskeletal pathology mechanisms. For additional information, please contact Ascent Research.
