The KIF5B Knockout Jurkat Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of Homo sapiens Jurkat T-lymphocyte leukemia cells with targeted disruption of the KIF5B gene. This polyclonal knockout product provides a heterogeneous loss-of-function model for studying kinesin-1 heavy chain function in a widely used immortalized T-cell line. The gene-edited pool retains the parental Jurkat background while offering a versatile tool for examining the consequences of KIF5B deficiency in intracellular transport, signaling, and cellular processes relevant to T-cell biology and malignancy.
The Jurkat cell line, established from the peripheral blood of a 14-year-old male with acute T-cell lymphoblastic leukemia, serves as a well-characterized model for T-lymphocyte activation, signaling, and apoptosis. These suspension cells express T-cell receptors (TCRs) and key signaling molecules, enabling robust studies of TCR engagement, co-stimulation, and downstream cascades. The immortalized nature and ease of manipulation make Jurkat cells a standard platform for investigating T-cell leukemia pathobiology and for screening immunomodulatory compounds.
KIF5B encodes the heavy chain of kinesin-1, a microtubule motor protein responsible for anterograde transport of organelles, vesicles, and protein complexes along microtubules. In Jurkat cells, KIF5B interacts with kinesin light chain (KLC1), JIP1/MAPK8IP1, JIP3/MAPK8IP3, GRB2 adaptor, huntingtin, 14-3-3 proteins, and the dynein motor complex. Its motor activity is regulated by upstream kinases including GSK3??, CDK1, and kinases downstream of TCR activation and CD28 costimulation, such as PI3K/AKT and MAP kinases. KIF5B-mediated transport directly influences mitochondrial trafficking, lysosomal positioning, autophagosome-lysosome fusion, immune synapse assembly, and mitotic spindle organization, while also scaffolding the transport of MEK-ERK signaling modules.
Disruption of KIF5B in Jurkat cells abolishes kinesin-1 motor function, impairing the coordinated transport of mitochondria, lysosomes, and signaling complexes along microtubules. This defect disrupts critical T-cell functions: TCR signaling is attenuated due to failed delivery of JIP?Cscaffolded MAP kinase modules to the immune synapse; mitochondrial redistribution, required for sustained activation, is blocked; and mitotic progression is compromised by improper spindle assembly. Consequently, the knockout cells exhibit altered T-cell activation marker expression, dysregulated apoptosis, and defective cell cycle dynamics, making them a potent model for dissecting the role of intracellular transport in T-cell leukemia and immune function.
Researchers can employ this polyclonal knockout model in a variety of advanced applications, including high-content immunofluorescence microscopy to quantify mitochondrial and lysosomal distribution, live-cell imaging of organelle motility, phospho-protein analysis of TCR-proximal kinases (e.g., pZAP70, pERK), flow cytometric assessment of activation markers CD69 and CD25, and cell cycle or apoptosis assays. The cells are suitable for RNAi or inhibitor screening aimed at kinesin-1 regulators and for studies of KIF5B-related transport defects in lymphoid malignancies. For further information, please contact Ascent Research.
