The KLHL7 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated through targeted disruption of the KLHL7 gene in the Jurkat T-lymphoblast cell line. This polyclonal knockout product provides a loss-of-function model with a heterogeneous pool of edited cells, enabling robust functional studies without the limitations of single-cell clonal selection. The cells retain the immortalized T-lymphocyte characteristics of the parental Jurkat line while lacking functional KLHL7 protein, making them suitable for investigating the role of KLHL7 in ubiquitin-mediated proteolysis and cell cycle regulation.
The host Jurkat cell line was originally derived from a patient with acute T-cell leukemia and serves as a well-established suspension cell model for T-cell receptor (TCR) signaling, T-cell activation, and leukemogenesis. These cells express components of the TCR/CD3 complex and exhibit characteristic T-lymphoblast morphology and growth properties. The suspension growth format facilitates high-throughput screening and scalable experimental workflows. As an immortalized T-cell line, Jurkat cells provide a reproducible background for studying gene function in lymphocyte biology and cancer-relevant pathways.
KLHL7 encodes a substrate-specific adaptor for the CUL3-RBX1 E3 ubiquitin ligase complex, which directs the ubiquitination and subsequent proteasomal degradation of target proteins. A key downstream target is Aurora B kinase, a critical mitotic regulator whose timely degradation is essential for proper chromosome segregation, cytokinesis, and mitotic exit. KLHL7 interacts directly with CUL3 and RBX1 to form the active E3 ligase, and this complex specifically recognizes Aurora B, conjugating ubiquitin chains that mark it for destruction by the 26S proteasome. Consequently, KLHL7 is a pivotal node connecting ubiquitin-proteasome system activity to mitotic progression and genome stability. Its function is regulated by upstream transcriptional mechanisms and post-translational modifications, and disruption of KLHL7-mediated proteostasis has been implicated in retinitis pigmentosa type 42 and possibly certain cancers.
In the Jurkat T-cell context, KLHL7 knockout provides a unique tool to dissect the crosstalk between ubiquitin-dependent protein degradation and T-cell physiology. Loss of KLHL7 is expected to elevate Aurora B levels, leading to mitotic defects, altered cell cycle profiles, and potential genomic instability??all of which can be monitored in this leukemia-derived background. This model is particularly relevant for exploring how dysregulation of the CUL3-RBX1-KLHL7 axis contributes to malignant transformation and for testing therapeutic strategies that target the ubiquitin-proteasome system in leukemia. Moreover, because Jurkat cells retain many features of normal T-cell signaling, the knockout line can be used to study whether KLHL7 influences TCR-mediated activation, proliferation, or apoptosis, bridging ubiquitin biology and immunology.
Researchers can employ these polyclonal knockout cells in a wide range of experimental workflows. Ubiquitination assays and co-immunoprecipitation of CUL3 complexes can be performed to validate the loss of KLHL7-dependent ligase activity, while Western blotting enables monitoring of Aurora B accumulation. Cell cycle analysis by flow cytometry and immunofluorescence microscopy for mitotic markers (e.g., phospho-histone H3) reveal mitotic defects. For T-cell-focused studies, activation and apoptosis assays can be combined with RT-qPCR to assess downstream transcriptional changes. The line is also suitable for drug screening aimed at identifying modulators of ubiquitin pathway components or compounds that selectively target leukemia cells with compromised proteostatic networks. For further information regarding this product, please contact Ascent Research.
