The IFT20 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Jurkat T lymphocyte line. This loss-of-function model is designed to study intraflagellar transport protein 20 (IFT20) in a T cell context, providing a genetically disrupted background for investigating ciliary trafficking, immune synapse architecture, and T cell signaling without the biases of clonal selection.
The Jurkat host cell line is an immortalized human T lymphocyte line originally isolated from an acute T cell leukemia patient. Widely used as a model for T cell receptor (TCR) signaling and activation, Jurkat cells recapitulate early TCR signaling events and are amenable to high-throughput screening and mechanistic studies. In this knockout population, they offer a physiologically relevant platform for examining the non-ciliary, synapse-related functions of IFT20.
IFT20 is a core subunit of the intraflagellar transport complex B (IFT-B), interacting with IFT52, IFT57, IFT88, and the dynein-2 motor to mediate ciliary trafficking and Hedgehog (Hh) pathway regulation. Its expression is controlled by RFX transcription factors such as RFX3 and is influenced by TCR activation signals. By facilitating ciliary transport, IFT20 enables processing of Hh pathway components such as Smoothened (SMO) and GLI transcription factors. In T cells, upon TCR engagement, IFT20 is recruited to the immune synapse via interaction with GMAP210 (TRIP11) and participates in TCR/CD3 clustering. This synaptic localization promotes phosphorylation of the proximal kinases ZAP70 and the adaptor LAT, thereby coupling TCR stimulation to downstream signaling cascades.
As Jurkat cells do not typically form primary cilia, IFT20??s role at the immune synapse becomes predominant. IFT20 gene disruption in these polyclonal cells enables dissection of its contribution to TCR microcluster organization, signal initiation, and T cell activation. The polyclonal population models heterogeneous IFT20 loss, allowing assessment of signaling variability within a cell population??valuable for studies of immune cell signaling robustness.
These IFT20 knockout Jurkat polyclonal cells are well-suited for immunofluorescence microscopy to examine immune synapse architecture and TCR clustering, flow cytometry to measure T cell activation markers including CD69 and phospho-ZAP70, and biochemical analyses such as western blotting and co-immunoprecipitation to assess IFT complex integrity and proximal TCR signaling. The knockout model further supports high-throughput screening for small-molecule modulators of TCR signaling and drug sensitivity testing using antileukemic agents like imatinib or dasatinib. For more information, please contact Ascent Research.
