The ANO10 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population originating from the human Jurkat T lymphocyte line, created to disrupt the ANO10 gene and eliminate its chloride channel and scramblase functions. This genetically heterogeneous pool provides a robust loss-of-function platform for dissecting ANO10-mediated ion and lipid dynamics in T-cell receptor signaling contexts. These cells serve as a powerful tool for exploring the downstream consequences of ANO10 ablation in immune cell signaling.
The Jurkat cell line, an immortalized human T lymphocyte derived from acute T cell leukemia, is a canonical model for T-cell receptor signaling, calcium mobilization, and immune response studies. Its acute leukemia origin endows it with robust proliferative capacity and well-defined signaling cascades, including stimulus-induced intracellular Ca2+ elevation that directly activates ANO10. The Jurkat background thus permits investigation of how ANO10-mediated chloride flux and phospholipid scrambling intersect with T-cell activation pathways, cytokine production, and membrane dynamics.
ANO10 encodes a dual-function protein that acts both as a calcium-activated chloride channel and a phospholipid scramblase. Activation is triggered by intracellular Ca2+ elevation, often downstream of GPCR-mediated IP3 production and CaMKII-mediated phosphorylation, and requires PIP2 binding. Upon activation, ANO10 binds calmodulin and facilitates chloride efflux and phosphatidylserine externalization, modulating membrane potential and cell volume while altering membrane lipid asymmetry. ERM proteins??ezrin, radixin, and moesin??interact with ANO10, anchoring it to the cytoskeleton and regulating its spatial activity. Collectively, these interactions position ANO10 as an integrator of calcium signals into ion and lipid fluxes critical for cellular homeostasis.
Disruption of ANO10 in Jurkat cells using CRISPR/Cas9 generates a polyclonal population lacking functional channel and scramblase activities, offering a direct means to assess ANO10-dependent processes. Loss of ANO10 abrogates Ca2+-induced phosphatidylserine exposure, quantifiable by Annexin V flow cytometry, and eliminates Ca2+-activated chloride currents, measurable by chloride-sensitive fluorescent dyes or patch clamp electrophysiology. This knockout model also enables examination of associated changes in cell volume regulation and membrane potential, which are essential for normal T-cell responses such as immunological synapse formation and activation-induced cell swelling.
Research applications for these polyclonal knockout cells span functional studies of calcium-activated chloride channels, dissection of phospholipid signaling networks in T cells, drug screening for ANO10 inhibitors or activators, and exploration of pathogenic mechanisms underlying spinocerebellar ataxia type 10 (SCAR10) and cerebellar atrophy. Typical experiments integrate calcium flux assays, phospholipid scrambling probes, and immunoblotting for ANO10 to validate knockdown and downstream signaling effects. For additional technical information or to inquire about custom reagents, please contact Ascent Research.
