ARID4A Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population derived from the human HT29 colorectal adenocarcinoma cell line. This product provides a loss-of-function model for studying the ARID4A tumor suppressor in an epithelial cancer context. The heterogeneous polyclonal format captures a range of disruption events, supporting robust functional interrogation without the constraints of clonal selection.
HT29 cells originate from a primary colorectal adenocarcinoma and serve as a well-established model for intestinal epithelial biology, drug absorption, and colorectal cancer research. These cells exhibit typical epithelial morphology and retain signaling pathways relevant to oncogenesis, including Wnt and MAPK cascades. Their use in ARID4A disruption provides a clinically pertinent system to examine gene function within the colorectal tumor microenvironment.
ARID4A is a transcriptional corepressor that interacts with RB1 and E2F1 to silence E2F target genes, thereby restraining cell cycle progression. It forms complexes with SIN3A, HDAC1, and SWI/SNF chromatin remodeling factors to coordinate epigenetic regulation. ARID4A is transcriptionally regulated by E2F factors and functionally linked to TP53 signaling, modulating downstream effectors such as CCNE1, CDKN1A, and BCL2 family members. Its loss disrupts repressive control of the RB/E2F axis and p53-dependent checkpoints, promoting unscheduled proliferation.
In HT29 cells, ARID4A knockout recapitulates a common colorectal cancer lesion, where loss of this tumor suppressor leads to deregulated proliferation, enhanced migratory capacity, and altered drug sensitivity. The HT29 background provides a relevant platform to dissect ARID4A??s role in Wnt-driven and p53-modulated colorectal carcinogenesis. This model thus enables investigation of tumor suppressive mechanisms and their consequences on epithelial homeostasis.
Researchers can employ these polyclonal knockout cells in a variety of assays: Western blotting and co-immunoprecipitation to confirm ARID4A absence and its interaction with RB1; RT-qPCR profiling of E2F target genes; chromatin immunoprecipitation to assess RB1/E2F1 occupancy; proliferation assays (MTT, BrdU) and Transwell migration assays; and apoptosis analysis via Annexin V staining. RNA-sequencing can reveal transcriptome-wide changes, while combinatorial drug screens can test ARID4A-dependent sensitivities. For additional technical details, please contact Ascent Research.
