ASGR1 Knockout HepG2 Cell Line

ASGR1 Knockout HepG2 Cell Line is a HepG2 liver cancer cell line in which ASGR1 (Asialoglycoprotein Receptor 1), the functional subunit of the asialoglycoprotein receptor (ASGPR) complex, is no longer expressed due to CRISPR/Cas9 genome editing. This cell line was generated by electroporation of HepG2 cells with ribonucleoprotein complexes of Cas9 with an sgRNA targeting ASGR1. This cell line has been validated by genomic sequencing and flow cytometry. The asialoglycoprotein receptor (ASGPR) complex is primarily expressed on the surface of hepatocytes, and is composed of two subunits, asialoglycoprotein receptor 1 and 2 (ASGR1 and ASGR2). ASGPR binds glycoproteins with a terminal galactose (Gal) or N-acetylgalactosamine (GalNAc), directing them to endocytosis and degradation in the lysosome. ASGR1 is the functional subunit of the ASGPR complex that binds to ligands and mediates endocytosis. Because of its liver localization, endocytosis activity, and efficient recycling back to the plasma membrane, ASGR1 has been used for liver-targeted drug delivery. Givosiran (Givlaari), a treatment for a genetic liver disease, was the first GalNAc-conjugated siRNA to be approved by the FDA. Several other GalNAc-conjugated siRNAs have since been approved or are undergoing clinical trials for the treatment of hereditary liver diseases, hepatitis B virus, or metabolic disorders. For example, Inclisiran (Leqvio) targets PCSK9 (Proprotein convertase subtilisin/kexin type 9) and is under investigation for the treatment of hypercholesterolemia. ASGR1 deficiency itself is associated with increased cholesterol efflux, decreased lipid levels in blood plasma and the liver, and decreased susceptibility to cardiovascular disease, suggesting that ASGR1 may be itself a promising therapeutic target. Finally, ASGR1 is being explored as a receptor for lysosome-targeting chimeras (LYTACs), which use GalNAc to direct extracellular or membrane-bound proteins to ASGR1 for endocytosis and degradation in the lysosome.