The ABCC4 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line, designed for targeted disruption of the ABCC4 gene. This product provides a loss-of-function model for investigating ABCC4-mediated transport and signaling in an epithelial intestinal cancer context. The polyclonal population consists of a heterogeneous pool of cells with CRISPR/Cas9-mediated ABCC4 gene disruption, enabling robust assessment of gene function without clonal selection biases.
The HT29 host cell line is a widely used human colorectal adenocarcinoma model of epithelial origin, serving as a representative system for studies of intestinal biology, colorectal cancer progression, and drug response. HT29 cells exhibit properties of absorptive and secretory intestinal epithelial cells, making them suitable for transport and signaling studies. Their well-characterized genetic background and established use in cancer research provide a relevant platform for ABCC4 knockout investigations.
ABCC4 encodes an ATP-binding cassette transporter that mediates ATP-dependent efflux of cyclic nucleotides (cAMP and cGMP), drugs, and organic anions. Upstream regulators include NRF2, HIF-1??, MAP kinases, and PXR, while downstream targets involve intracellular cAMP/cGMP levels, PKA, CREB, and drug accumulation. The transporter interacts with scaffold proteins such as NHERF1, PDZK1, and EBP50, which localize it to membrane domains. By exporting cAMP and cGMP, ABCC4 modulates signaling through PKA and CREB, as well as prostaglandin synthesis via the COX-2/PGE2 pathway. Knockout of ABCC4 disrupts these signaling cascades and alters cellular responses to transported substrates.
In HT29 colorectal adenocarcinoma cells, ABCC4 knockout provides a model for investigating chemoresistance, as many anticancer drugs are substrates for ABCC4-mediated efflux. The loss of ABCC4 function elevates intracellular cyclic nucleotide levels, which can enhance PKA/CREB signaling and influence cell proliferation, differentiation, and apoptosis. This system allows dissection of how ABCC4 contributes to drug sensitivity and inflammation-associated pathways in an intestinal epithelial context. Moreover, it enables study of compensatory transporter upregulation and signaling rewiring that may occur upon ABCC4 disruption.
Applications include drug resistance assays (e.g., cell viability with methotrexate or 6-mercaptopurine), transporter functional studies using fluorescent substrates like calcein-AM, intracellular cAMP ELISA, and PKA activity assays. Additional experiments such as wound healing and colony formation can assess migratory and clonogenic changes. RT-qPCR and Western blotting confirm knockout. This model thus supports research in cancer biology, drug transport, and signaling. For more information, please contact Ascent Research.
