The AGA Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in the HAP1 line, designed to disrupt the AGA gene encoding aspartylglucosaminidase. This heterogeneous pool ensures robust loss of gene function across the culture, providing a reliable model without the constraints of single-cell clonal selection. The product is suited for investigating lysosomal glycoprotein catabolism and aspartylglucosaminuria pathology.
HAP1 is a human near-haploid adherent cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploidy permits efficient gene knockout, as a single editing event eliminates the sole allele, generating clear loss-of-function phenotypes. The line offers a stable karyotype and routine culture, making it a workhorse for functional genomics, cancer research, and organelle biology studies, while its leukemic origin adds relevance for hematological disease modeling.
Aspartylglucosaminidase, the product of AGA, catalyzes the cleavage of the N-glycosidic bond linking asparagine to N-acetylglucosamine in glycoproteins within lysosomes, initiating the release of aspartate and free oligosaccharides. This enzyme acts in concert with lysosomal proteases such as cathepsin D and other glycosidases including ??-mannosidase, ??-hexosaminidase, ??-galactosidase, and sialidase to ensure complete glycoprotein degradation. AGA transcription is regulated by the MiT/TFE family??TFEB, MITF, and TFE3??in response to lysosomal stress, placing it within a broader network of lysosomal biogenesis control. Consequently, AGA knockout disrupts this catabolic cascade, causing accumulation of glycoasparagines characteristic of aspartylglucosaminuria.
In haploid HAP1 cells, AGA inactivation directly models the metabolic defect of aspartylglucosaminuria, with polyclonal knockout ensuring population-level deletion and eliminating clonal artifacts. The model enables study of lysosomal storage pathology, including impaired autophagy and lysosomal enlargement, and facilitates rescue experiments due to the absence of wild-type alleles. The leukemic background also allows exploration of lysosomal function in cancer cell physiology.
Applications include drug screening for enzyme replacement and small-molecule therapies, validation of CRISPR targeting, and dissection of glycoprotein degradation pathways. Typical assays comprise RT-qPCR and western blotting for expression analysis, enzymatic activity measurements, immunofluorescence and LysoTracker staining for lysosomal changes, and mass spectrometry for glycoasparagine profiling. For further information, please contact Ascent Research.
