This product comprises a polyclonal population of Jurkat cells in which the ANO6 gene has been disrupted using CRISPR/Cas9 gene editing, resulting in loss of scramblase and ion channel functions. The polyclonal format retains a heterogeneous mixture of edited alleles, minimizing clonal selection artifacts and providing a robust platform for functional assays. These cells are supplied as a ready-to-use research tool, enabling immediate investigation of ANO6-dependent processes in a human T lymphoblast background. Additionally, this knockout model serves as an ideal control for studies requiring abrogation of calcium-activated phospholipid scrambling, such as annexin V binding assays or phosphatidylserine externalization measurements.
Jurkat cells are a human immortalized T lymphoblast line derived from an acute T cell leukemia patient. They serve as a universal model for T lymphocyte biology, expressing CD3 and CD28 and responding to stimulation with calcium flux and cytokine secretion. Jurkat cells are widely used for studying signal transduction, apoptosis, and immune modulation. Their well-characterized signaling networks and genetic tractability make them ideal for CRISPR/Cas9-mediated gene disruption, enabling precise dissection of gene function in a T cell context. Additionally, these cells can be stimulated through their T cell receptor to activate downstream pathways, facilitating analysis of ANO6 in T cell activation contexts.
ANO6 (TMEM16F) is a dual-function transmembrane protein that acts as a calcium-activated phospholipid scramblase and a non-selective ion channel. It mediates phosphatidylserine translocation from the inner to the outer plasma membrane leaflet, a process critical for blood coagulation factor activation and clearance of apoptotic cells. Upstream regulators include intracellular calcium and caspases, while downstream effects encompass phosphatidylserine externalization, microvesicle shedding, and coagulation cascade engagement. ANO6 forms homodimers and associates with the actin cytoskeleton, integrating calcium signals with membrane asymmetry. Thus, ANO6 bridges calcium signaling, lipid remodeling, and hemostatic/apoptotic pathways.
Disruption of ANO6 in Jurkat cells eliminates calcium-dependent scramblase activity, enabling detailed investigation of phosphatidylserine exposure in a lymphocyte context. As Jurkat cells undergo apoptosis-driven externalization, this knockout population serves as a negative control for studying apoptotic cell clearance and microvesicle formation. Additionally, it illuminates potential scramblase roles in T cell fusion and leukemic cell biology. The polyclonal nature reduces clonal phenotypic variation, ensuring consistent results in functional assays such as annexin V labeling, while preserving a loss-of-function background for pathway analysis. Moreover, this model can be used to assess the contribution of ANO6 to T cell receptor-induced signaling and lipid raft dynamics.
This polyclonal ANO6 knockout cell population supports a spectrum of research areas: Scott syndrome modeling, platelet function analysis (using Jurkat as a surrogate), and phosphatidylserine-dependent signaling studies. In cancer biology, it permits exploration of scramblase contributions to leukemic cell survival and immune evasion. Compatible assays include flow cytometric annexin V binding, calcium flux measurements, phosphatidylserine externalization quantification, western blot analysis, and RT-qPCR. These cells are also suitable for drug screening applications targeting scramblase activity or for studying the role of ANO6 in microvesicle-mediated intercellular communication. For further information or to discuss bulk orders, please contact Ascent Research.
