The ALG3 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Jurkat cells in which the ALG3 gene has been disrupted, resulting in loss of ALG3 alpha-1,3-mannosyltransferase function. This knockout model provides a valuable tool for investigating the roles of dolichol-linked oligosaccharide synthesis and protein N-glycosylation in T lymphocyte biology. The polyclonal nature of the product ensures representation of diverse editing events within the population, offering a robust system for functional studies without clonal selection biases.
The parental Jurkat cell line is a widely used human T lymphocyte model derived from an acute T cell leukemia patient. Jurkat cells are instrumental in dissecting T cell receptor (TCR) signaling pathways, activation mechanisms, and apoptosis regulation. Their well-characterized signal transduction network makes them an ideal host for studying how post-translational modifications, particularly glycosylation, influence immune cell function and fate decisions.
ALG3 encodes an alpha-1,3-mannosyltransferase that catalyzes the addition of the sixth mannose residue to the growing dolichol-linked oligosaccharide precursor Man5GlcNAc2-PP-dolichol, generating Man6GlcNAc2-PP-dolichol. This step is essential in the biosynthetic pathway of N-glycans. ALG3 function is integrated into the endoplasmic reticulum (ER) glycosylation machinery, acting downstream of dolichol phosphate mannose synthesis by DPM1, and interacts with other ALG mannosyltransferases such as ALG1, ALG2, ALG6, and ALG8. Its product is further processed by ALG9 and ALG12 before transfer to nascent proteins by the oligosaccharyltransferase complex (including STT3A/B). Consequently, ALG3 disruption leads to defective N-glycosylation of a broad range of proteins, including TCR subunits and cytokine receptors, triggering ER stress and activating the unfolded protein response (UPR) via sensors IRE1, PERK, and ATF6, and downstream transcription factors XBP1 and ATF6.
In Jurkat T cells, impaired N-glycosylation due to ALG3 knockout profoundly impacts cell surface receptor expression, stability, and signaling competence. This system models the molecular pathology of congenital disorder of glycosylation type Id (ALG3-CDG) and allows investigation of how glycosylation defects alter T cell activation, proliferation, and apoptosis. The interplay between ER stress and immune signaling pathways can be systematically explored, providing insights into the cellular consequences of glycosylation insufficiency.
Researchers can utilize ALG3 Knockout Jurkat Polyclonal Cells to perform Western blot analysis of glycosylation-sensitive markers such as TCR???? and ICAM-1, lectin blotting with ConA or PNA, and RT-qPCR quantification of UPR targets like BiP and CHOP. Flow cytometry enables assessment of surface glycoprotein levels, while Annexin V staining and proliferation assays evaluate functional outcomes. These cells are also suitable for pharmacological studies involving ER stress inducers (e.g., tunicamycin) or potential therapeutic compounds. For additional information on using this model in your studies, please contact Ascent Research.
