The ABCC5 Knockout HT29 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal population with disruption of the ABCC5 gene, generating a loss-of-function model of the multidrug resistance protein 5 (MRP5). This heterogeneous mixture of edited cells provides a robust tool for studying ABCC5 function without clonal isolation.
The HT29 cell line is a human colorectal adenocarcinoma isolated from a 44-year-old female. It models intestinal epithelium, forming tight junctions and microvilli, and harbors oncogenic mutations in APC, TP53, and KRAS, which are common in colorectal cancer. These features make HT29 a relevant system for investigating tumor biology and drug responses.
ABCC5 encodes MRP5, an ATP-binding cassette transporter that effluxes cyclic nucleotides (cAMP, cGMP) and nucleotide analog drugs such as 5-fluorouracil and methotrexate. Regulated by NRF2 and ??-catenin, and induced by xenobiotics and oxidative stress, MRP5 interacts with PDZ-domain proteins like NHERF1 and cytoskeletal linker ezrin. By exporting cyclic nucleotides, MRP5 dampens downstream PKA and PKG signaling. Its knockout leads to intracellular accumulation of cAMP and cGMP, potentially enhancing cyclic nucleotide-dependent pathways and reversing drug resistance.
In the HT29 colorectal adenocarcinoma context, ABCC5 knockout enables dissection of MRP5-mediated drug resistance and cyclic nucleotide homeostasis. Overexpression of ABCC5 in colorectal cancer correlates with poor prognosis, making this model valuable for studying how transporter loss affects drug sensitivity and signaling. The interaction between ABCC5 and the Wnt/??-catenin pathway, disrupted by APC mutation in HT29, allows exploration of oncogenic and transport pathway crosstalk. Additionally, the model is applicable to inflammatory bowel disease research, given the role of cyclic nucleotide signaling in intestinal epithelial function.
This polyclonal knockout population supports diverse applications: cancer drug resistance studies, high-throughput drug screening, cyclic nucleotide signaling analysis, and transport assays. Representative techniques include western blotting, RT-qPCR, MTT viability assays, intracellular cAMP/cGMP ELISA, fluorescent efflux assays (e.g., calcein-AM), and apoptosis assays (Annexin V/PI). Researchers can screen for ABCC5 modulators or validate cGMP-PKG and cAMP-PKA pathway intermediates. For further information, contact Ascent Research.
