The IFT74 Knockout Jurkat Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout population of Jurkat human T lymphocytes with disruption of the IFT74 gene. This product provides a heterogeneous knockout model to investigate IFT74 loss-of-function without clonal selection bias. The gene encodes a core intraflagellar transport protein, enabling studies of ciliary assembly and signaling in a T-cell context.
Jurkat cells are an immortalized T-lymphocyte line derived from a patient with acute T-cell leukemia, widely used to study T-cell receptor signaling, apoptosis, and immune function. Their robust proliferation and genetic tractability make them suitable for CRISPR/Cas9-mediated gene editing. In this polyclonal format, they allow examination of IFT74??s role in both ciliary and non-ciliary processes within an immune-competent cellular background.
IFT74 is a critical component of the intraflagellar transport complex B (IFT-B), essential for retrograde trafficking along ciliary microtubules. It interacts with IFT-B subunits IFT81, IFT88, IFT52, IFT27, IFT20, and the motor proteins kinesin-2 and dynein-2. Transcription is controlled by RFX factors, FOXJ1, and Notch signaling. Disruption of IFT74 impairs IFT-B assembly, blocking ciliogenesis, and attenuates Hedgehog signaling by reducing Smoothened-dependent activation of Gli transcription factors. Cross-talk with Wnt pathways may also be affected.
The Jurkat background offers a unique platform to dissect IFT74 function in immune cells. Although T lymphocytes are not constitutively ciliated, they can form primary cilia under certain conditions, and ciliary proteins may have non-ciliary roles. This knockout model allows assessment of cilia-dependent functions like ciliogenesis and Hedgehog signal transduction, alongside potential cilia-independent roles in T-cell proliferation, apoptosis, or cytokine response. The polyclonal nature reduces clonal variation, providing a more representative population for functional assays.
Typical applications include ciliogenesis assays under serum starvation, validation of knockout efficiency by Western blot and RT-qPCR, and co-immunoprecipitation to evaluate IFT-B complex integrity. Immunofluorescence for ciliary markers (acetylated tubulin), flow cytometry for cell cycle analysis, and drug screening for ciliopathy targets are also feasible. This product serves as a robust tool for studying intraflagellar transport and translational research into skeletal ciliopathies such as short-rib thoracic dysplasia. For further information, please contact Ascent Research.
