The EBP Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal carcinoma cell line HCT 116. This product offers a targeted loss-of-function model for the EBP gene, achieved through CRISPR/Cas9-mediated gene disruption, and is supplied as a heterogeneous pool of edited cells to facilitate studies requiring diverse mutational outcomes.
The HCT 116 host cell line is a well-characterized model of colorectal carcinoma, exhibiting microsatellite instability-high (MSI-H) status, a KRAS G13D mutation, a CTNNB1 mutation, and wild-type TP53. These epithelial cells are widely employed in cancer research for investigating oncogenic signaling, metastatic mechanisms, and drug sensitivity, making them a suitable background for dissecting the role of cholesterol metabolism in colorectal tumor biology.
EBP encodes the sterol delta8-isomerase, an endoplasmic reticulum enzyme that catalyzes the conversion of delta8-cholesterol to delta7-cholesterol in the final stages of cholesterol biosynthesis. This reaction acts downstream of sterol regulatory transcription factors such as SREBP1 and SREBP2, which are activated by low intracellular cholesterol and insulin signaling. EBP functions in concert with SC5D (sterol C5-desaturase) and DHCR7 (7-dehydrocholesterol reductase) to produce cholesterol, essential for membrane structure and steroidogenesis. Disruption of EBP leads to accumulation of delta8-cholesterol and impaired generation of downstream products including lathosterol and 7-dehydrocholesterol, ultimately reducing cholesterol and steroid hormone synthesis.
In the context of HCT 116 cells, EBP knockout is expected to perturb cholesterol homeostasis and lipid raft integrity, potentially altering signaling pathways driven by oncogenic KRAS and CTNNB1 mutations. The model provides a platform to examine how disrupted sterol biosynthesis impacts colorectal carcinoma cell proliferation, survival, and metastatic potential. Additionally, the polyclonal nature of the knockout population allows for the observation of phenotypic variability associated with different editing events, offering insights into the functional heterogeneity of cholesterol pathway dependencies in cancer.
Researchers can utilize these EBP knockout cells in a broad range of applications, including detailed cholesterol metabolism studies using lipidomics and cholesterol quantification assays such as Amplex Red or HPLC. The cells are suitable for drug screening of cholesterol pathway inhibitors like tamoxifen and U18666A, and for investigating mechanisms underlying cholesterol-related disorders such as Conradi-H??nermann syndrome. Functional assays such as Western blotting, RT-qPCR, RNA-seq, and flow cytometry for lipid raft markers can be employed to assess changes in sterol-responsive gene expression and protein localization. For further information, please contact Ascent Research.
