The ABCA1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the human ABCA1 gene in the HCT 116 colorectal carcinoma cell line. This polyclonal format provides a heterogeneous mix of gene-edited cells, enabling robust phenotypic screening without clonal isolation. The loss of ABCA1 protein function creates a powerful model for dissecting pathways of cholesterol efflux and high-density lipoprotein (HDL) biogenesis in a cancer-relevant epithelial context.
The host HCT 116 cell line is an epithelial colorectal carcinoma line with wild-type p53 and proficient mismatch repair, extensively used in cancer biology for studying tumorigenesis, apoptosis, and drug responses. Its well-defined genetic background and ease of culture make it a preferred system for genetic loss-of-function studies, allowing researchers to examine the impact of ABCA1 deletion on lipid metabolism within a malignant setting.
ABCA1 encodes a plasma membrane transporter that mediates the rate-limiting step in HDL formation by effluxing cholesterol and phospholipids to lipid-poor apolipoprotein A-I (apoA-I). Its transcription is activated by liver X receptor alpha/beta (LXR??/??) and retinoid X receptor (RXR) heterodimers upon binding oxysterols, PPAR?? agonists, or synthetic LXR agonists like T0901317. ABCA1 interacts directly with apoA-I, Cdc42, and ??1-syntrophin, and participates in reverse cholesterol transport alongside ABCG1, scavenger receptor class B type I (SR-BI), lecithin-cholesterol acyltransferase (LCAT), and cholesteryl ester transfer protein (CETP). Downstream, this cascade yields mature HDL particles and regulates cellular cholesterol homeostasis.
In HCT 116 cells, ABCA1 knockout uncouples cholesterol efflux from lipoprotein biogenesis, enabling dissection of lipid metabolic reprogramming common in colorectal cancers. Cancer cells often upregulate cholesterol uptake and storage, and loss of ABCA1 can shift intracellular cholesterol distribution, affecting lipid raft composition, signal transduction, and LXR target gene expression. This model thus provides a platform to study how cholesterol flux influences cancer cell proliferation, migration, and sensitivity to chemotherapeutics or targeted agents.
These knockout cells are amenable to cholesterol efflux assays using fluorescent or radiolabeled cholesterol, HDL particle reconstitution with recombinant apoA-I, Western blotting and qPCR for ABCA1 and LXR-regulated genes (e.g., ABCG1, SREBF1), confocal imaging of intracellular lipid droplets, and flow cytometry for cell surface lipid markers. They support drug discovery for atherosclerosis, Tangier disease, and Alzheimer’s disease, as well as basic research into lipid-related cancer mechanisms. For further inquiries, please contact Ascent Research.
