The ASGR1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the human colorectal carcinoma HCT 116 cell line, designed to disrupt the ASGR1 gene encoding the major asialoglycoprotein receptor subunit. This polyclonal format comprises a heterogeneous pool of cells with distinct editing events at the ASGR1 locus, providing a robust loss-of-function model without the need for clonal selection. It is ideal for high-throughput screening and functional studies of receptor-mediated endocytosis.
The HCT 116 host cell line is a well-characterized model of colorectal carcinoma harboring a KRAS G13D mutation, MLH1 deficiency, and high microsatellite instability (MSI-H). These genetic alterations drive constitutive MAPK signaling and defective DNA mismatch repair, contributing to a tumorigenic phenotype. The epithelial adherent morphology facilitates standard cell culture, transfection, and imaging-based assays. This background offers a relevant isogenic platform to study interactions between oncogenic signaling and glycoprotein homeostasis.
ASGR1, in complex with ASGR2, forms the hepatic asialoglycoprotein receptor that mediates endocytosis of desialylated glycoproteins. Expression is transcriptionally regulated by HNF1A and HNF4A and induced by glucocorticoids. Upon ligand binding, ASGR1 recruits clathrin and adaptor protein AP2, initiating clathrin-dependent endocytosis. Dynamin facilitates vesicle scission, leading to lysosomal degradation by lysosomal hydrolases. This clearance is critical for removal of apoptotic cells and immune tolerance; knockout abrogates these functions, potentially disrupting glycoprotein homeostasis.
Within the HCT 116 colorectal carcinoma context, ASGR1 knockout introduces a distinct perturbation. The existing KRAS G13D and MLH1 defects already promote apoptosis resistance, and loss of asialoglycoprotein clearance may further disrupt apoptotic cell removal and immune surveillance. This model enables dissection of glycoprotein trafficking and its impact on tumor biology. Since ASGR1 is canonically studied in hepatocytes, these cells allow investigation of receptor function in a non-hepatic cancer line, facilitating broader studies of ligand specificity and endocytic trafficking.
These polyclonal knockout cells are applicable to diverse assays, including western blotting and RT-qPCR for knockout confirmation, immunofluorescence and flow cytometry for expression and endocytosis monitoring, and functional endocytosis assays using fluorescent asialofetuin. Apoptosis and proliferation assays can assess consequences of impaired clearance. Research areas include glycoprotein homeostasis, ASGR1 function in colorectal cancer, drug delivery targeting via the asialoglycoprotein receptor, and immune tolerance modulation. For further details, contact Ascent Research.
