The KDSR Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in the Jurkat T?lymphoblast line, featuring targeted disruption of the KDSR gene. This product provides a genetically heterogeneous pool of cells with KDSR loss?of?function, generated without clonal isolation, enabling robust population?level studies of sphingolipid metabolism. The polyclonal format avoids the artifacts of single?cell selection while preserving the functional relevance of CRISPR/Cas9?mediated gene disruption, making it suited for applications requiring biological replicates and average phenotypic responses.
Jurkat cells were originally derived from a 14?year?old male with acute lymphoblastic leukemia and are widely employed as a model for human T?cell receptor (TCR) signaling, cytokine production, and immune activation. They constitutively express CD3, CD4, and the IL?2 receptor, and respond to phorbol esters and lectins with robust interleukin?2 secretion. The leukemic origin endows these cells with a well?characterized signaling network that is particularly amenable to studies of apoptosis and proliferation, while their suspension?growth characteristics facilitate high?throughput and multi?omic experimental workflows.
KDSR encodes 3?ketodihydrosphingosine reductase, which catalyzes the NADPH?dependent reduction of 3?ketodihydrosphingosine to sphinganine, the penultimate step in de novo ceramide biosynthesis. This reaction lies downstream of the serine palmitoyltransferase (SPT) complex and upstream of ceramide synthase and dihydroceramide desaturase. The enzyme is transcriptionally regulated by SREBP?1, LXR, and PPAR??, and its product sphinganine is a direct precursor of ceramide, sphingosine?1?phosphate, and other sphingolipids. KDSR activity is therefore a critical node controlling the balance between pro?apoptotic ceramide and pro?survival sphingosine?1?phosphate signals, with direct implications for cellular fate decisions.
In Jurkat T cells, sphingolipid metabolism modulates TCR?dependent signaling, Fas?mediated apoptosis, and IL?2 production. Disruption of KDSR depletes sphinganine and ceramide pools, leading to altered lipid raft composition, impaired caspase?8 activation, and attenuated death?receptor?induced apoptosis. This perturbation recapitulates key aspects of sphingolipid?related disorders including follicular lymphoma, where KDSR is linked to the FVT?1 locus, and neurodegenerative sphingolipidoses. The Jurkat background offers a tractable platform to dissect how KDSR?dependent lipid changes affect kinase cascades, mitochondrial integrity, and nuclear factor???B activation downstream of the TCR.
Typical applications integrate this KDSR?knockout model with LC?MS?based sphingolipid profiling, ceramide quantification, Annexin V/PI apoptosis assays, cleaved caspase?3 Western blotting, and RT?qPCR for sphingolipid?enzyme expression. Researchers also employ these cells in proliferation assays and chemical screens for sphingolipid?pathway modulators, including SPT inhibitors and ceramide synthase activators. The polyclonal population is suitable for studying variable penetrance of sphingolipid?dependent phenotypes and for co?culture experiments investigating T?cell?mediated cytotoxicity. For further information and custom applications, please contact Ascent Research.
