The ARL8B Knockout Jurkat Polyclonal Cells offered by Ascent Research provide a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T-lymphocyte cell line. This product features targeted disruption of the ARL8B gene, resulting in a loss-of-function model for studying the role of this small GTPase in lysosomal trafficking and cellular positioning pathways. The polyclonal format avoids clonal selection artifacts and maintains population-level heterogeneity, making it suitable for bulk functional assays and screening applications.
Jurkat cells are a widely used immortalized T-cell line originally isolated from the peripheral blood of a 14-year-old male with acute T-cell leukemia. They serve as a well-characterized in vitro model for T-cell signaling, activation, and apoptosis. Their robust growth, facile genetic manipulation, and relevance to human immunology make them ideal for exploring molecular mechanisms underlying T-cell effector functions, including cytokine secretion and cytotoxic activity.
ARL8B (ADP-ribosylation factor-like 8B) is a small GTPase associated with late endosomes and lysosomes, functioning as a key regulator of lysosome positioning via microtubule-mediated anterograde transport. The BORC complex, containing BORCS5 and BORCS6, activates ARL8B on lysosomal membranes by promoting nucleotide exchange. Active ARL8B recruits the adaptor SKIP (PLEKHM2), which links the GTPase to plus-end-directed kinesin motors KIF5B and KIF1B??, driving peripheral lysosome movement. In T lymphocytes, ARL8B-dependent trafficking is critical for concentrating lytic granules at the immunological synapse during cytotoxicity. Additionally, ARL8B participates in autophagosome-lysosome fusion and is regulated by mTORC1 and TFEB, situating it at a central hub coordinating nutrient sensing and organelle dynamics.
In Jurkat T cells, ARL8B-mediated positioning of lysosomes and lytic granules directly impacts immune synapse formation and granule exocytosis. Disruption of ARL8B allows dissection of spatial and temporal requirements for lysosome trafficking during T-cell activation and target cell killing. The knockout phenotype includes impaired lytic granule polarization, reduced CD107a exposure, and diminished cytotoxic capacity. These deficits make the polyclonal knockout cells a valuable tool for elucidating molecular requirements for immune surveillance and for identifying targets to modulate T-cell responses in cancer immunotherapy or autoimmunity.
Key research applications of the ARL8B Knockout Jurkat Polyclonal Cells include real-time imaging of lysosome dynamics using fluorescent markers, quantification of degranulation via CD107a flow cytometry, and functional cytotoxicity assays. The model is also suited for biochemical studies such as co-immunoprecipitation to map ARL8B-dependent protein interaction networks, and for monitoring autophagic flux. Furthermore, these cells provide a platform for small-molecule screening aimed at lysosomal trafficking modulators in lysosomal storage disorders and cancer. For further scientific inquiries or technical assistance, please contact Ascent Research.
