The ABCB1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-engineered polyclonal knockout cell population in which the ABCB1 gene has been disrupted via targeted editing. This product comprises a mixed population of HCT 116 cells with heterogeneous loss-of-function mutations in ABCB1, enabling functional studies of P-glycoprotein (P-gp) without clonal selection. The polyclonal format captures genetic diversity across edited alleles, making it suitable for pooled analyses where population-level phenotypes are desired.
The HCT 116 host cell line is a widely used human colorectal carcinoma epithelial model. These cells exhibit high microsatellite instability (MSI-H) due to MLH1 deficiency and harbor a heterozygous KRAS G13D mutation, reflecting genetic alterations common in colorectal cancers. HCT 116 cells are adherent, maintain epithelial morphology, and are extensively characterized in cancer research for studying tumor biology, signal transduction, and drug response.
ABCB1 encodes P-glycoprotein, an ATP-binding cassette (ABC) transporter that acts as a broad-specificity efflux pump for xenobiotics and chemotherapeutic drugs. P-glycoprotein expression is transcriptionally regulated by NF-??B, p53, HIF-1??, PXR, CAR, and MAPK signaling cascades, which converge on the ABCB1 promoter. The transporter interacts with Caveolin-1 and cholesterol within membrane microdomains to modulate its efflux activity. Downstream, ABCB1-mediated efflux reduces intracellular drug accumulation and attenuates drug-induced apoptosis. This gene is central to multidrug resistance, often cooperating with ABCG2, and is embedded in PI3K/AKT pathways frequently activated in cancer.
In the HCT 116 background, ABCB1 knockout provides a powerful system for dissecting drug resistance mechanisms in colorectal cancer. The MLH1-deficient, MSI-H status and KRAS G13D mutation create a context where multidrug resistance pathways may be upregulated, and loss of P-glycoprotein helps isolate its contribution to reduced drug sensitivity. Researchers can compare drug efflux rates, assess chemosensitivity, and explore interactions between ABCB1 and other resistance factors. The polyclonal population enables robust population-level assays, avoiding clonal bias and better representing heterogeneous tumor cell populations.
This product supports applications including multidrug resistance studies, ABC transporter functional analysis, drug screening, and chemosensitivity assays. Representative experiments include Western blotting and RT-qPCR for knockout validation, drug efflux assays with fluorescent substrates, flow cytometry for intracellular drug accumulation, and cytotoxicity testing. Immunofluorescence can assess P-glycoprotein localization. These cells provide a versatile platform for investigating drug resistance in colorectal cancer. For more information, please contact Ascent Research.
