The ASGR1 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1299 human non-small cell lung cancer line, genetically modified to disrupt expression of the ASGR1 gene. ASGR1 encodes the major subunit of the asialoglycoprotein receptor (ASGPR), a hepatic lectin responsible for mediating endocytosis of desialylated glycoproteins. This knockout reagent is provided as a heterogeneous pool of edited cells, enabling robust loss-of-function studies without clonal selection, and is suitable for investigating ASGPR-dependent processes in a lung adenocarcinoma background.
NCI-H1299 is a widely used epithelial cell line established from the lymph node metastasis of a human lung adenocarcinoma. It serves as a model for non-small cell lung cancer (NSCLC) biology, including mechanisms of invasion, metastasis, and therapeutic resistance. The parental line exhibits characteristic features of adenocarcinoma, such as KRAS mutation and epithelial morphology. By introducing ASGR1 knockout into this context, researchers can dissect the receptor??s role outside the canonical hepatic environment, potentially uncovering novel functions in cancer glycobiology.
ASGR1 forms a heterodimeric complex with ASGR2, operating within the clathrin-mediated endocytosis pathway. The receptor cycle involves assembly into clathrin-coated pits, interaction with adaptor protein complex 2 (AP2), and internalization into Rab5-positive early endosomes, ultimately delivering cargo to Lamp1-positive lysosomes for degradation. Upstream regulators of ASGR1 transcription include hepatic nuclear factors HNF1A and HNF4A, as well as STAT3, while epigenetic silencing has been observed in certain cancers. Downstream, loss of ASGR1 abolishes clearance of desialylated glycoproteins, leading to altered serum glycoprotein levels and potential modulation of the NF-??B pathway. Additionally, ASGR1 interacts with the hepatitis B virus surface antigen, linking it to viral entry mechanisms.
Ablation of ASGR1 in NCI-H1299 cells provides a unique platform for exploring the intersection of glycoprotein metabolism and lung adenocarcinoma pathobiology. The model is particularly relevant given the lung cancer environment, where aberrant glycosylation and altered endocytic trafficking are implicated in immune evasion and metastasis. By disrupting the asialoglycoprotein receptor, researchers can assess impacts on glycoprotein-mediated signaling, lysosomal degradation capacity, and innate immunity regulation??areas of growing interest in NSCLC. This knockout also facilitates study of the ASGPR as a potential target for glycoprotein-based drug delivery strategies in non-hepatic cancers.
This polyclonal knockout population is ideally suited for a range of experimental applications, including glycoprotein clearance assays using fluorescently labeled asialofetuin, characterization of ASGPR surface expression by flow cytometry, and transcriptional profiling via RNA-seq to identify downstream targets. Protein-level analysis can be performed through Western blotting and co-immunoprecipitation for ASGR1-ASGR2 interaction, while lysosomal degradation assays and cellular viability tests enable functional readouts. Researchers investigating ASGPR-mediated drug delivery, viral entry by hepatitis B virus, or glycobiology in cancer will find this model highly valuable. For further details, please contact Ascent Research.
