The ALG1L2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the ALG1L2 gene. Derived from HeLa cells, this knockout model enables functional studies of ALG1L2, a putative mannosyltransferase involved in N-glycosylation. The polyclonal format ensures a heterogeneous allele mixture representative of loss-of-function effects across the population.
HeLa cells, an HPV18-positive cervical adenocarcinoma line, are immortalized and epithelial, with p53 and Rb inactivated by viral oncoproteins E6 and E7. Their highly aneuploid genome and robust growth make them a standard host for cancer biology, gene function, and glycobiology research. The line??s well-documented characteristics facilitate integration into diverse experimental workflows.
ALG1L2 functions in the dolichol-linked oligosaccharide biosynthetic pathway, catalyzing mannose addition to the lipid-linked precursor for N-glycan transfer. It operates within a complex including DPM1, DPM2, and DPM3, and collaborates with ALG2 and ALG11. Transcription is driven by SP1 and E2F, while the unfolded protein response (UPR) sensors ATF6, IRE1, and PERK modulate its expression under ER stress. Downstream, defective ALG1L2 activity reduces mature dolichol-linked oligosaccharides, compromising glycosylation of integrins, growth factor receptors, and cell adhesion molecules.
In HeLa cells, disruption of ALG1L2 exacerbates glycosylation abnormalities inherent to the aneuploid and oncogene-driven background. This polyclonal knockout provides a tractable system to dissect how N-glycosylation defects influence cancer cell behavior, including adhesion, proliferation, and stress responses. The model also recapitulates features of congenital disorders of glycosylation (CDG), enabling mechanistic studies and therapeutic screening relevant to both CDG and aberrant cancer glycosylation.
Researchers can employ the ALG1L2 Knockout HeLa Polyclonal Cells in lectin blotting (ConA, PHA-L) to detect N-glycan alterations, western blotting for glycoprotein molecular weight shifts, and N-glycan profiling by mass spectrometry. ER stress can be monitored via immunofluorescence for UPR markers, while cell viability and tumorigenicity assays assess functional consequences of knockout. Dolichol-linked oligosaccharide analysis by HPLC allows quantification of lipid-linked intermediates. For additional information, contact Ascent Research.
