The DLGAP5 Knockout Jurkat Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the human DLGAP5 gene has been targeted for disruption. This product provides a heterogeneous pool of Jurkat T lymphocytes carrying diverse loss-of-function alleles, enabling pooled functional studies without clonal selection or single-cell isolation. It is designed for researchers investigating mitotic spindle regulation, chromosomal stability, and oncogenic signaling within a lymphoid background, offering a robust platform for both focused mechanistic assays and broader phenotypic screens. The polyclonal format reflects the complexity of genomic editing outcomes, facilitating the analysis of gene function across a population of edited cells.
The Jurkat host cell line is an immortalized human CD4+ T lymphocyte model originally derived from an acute T cell leukemia patient. Jurkat cells are extensively utilized to dissect T cell receptor signaling, apoptosis, and HIV pathogenesis, owing to their well-characterized intracellular pathways and ease of culture in suspension. Their transformed nature also makes them a relevant system for studying the molecular underpinnings of T cell malignancies, including proliferation defects and genomic instability. These features establish Jurkat cells as a suitable background for examining the mitotic roles of DLGAP5.
DLGAP5 encodes a mitotic spindle assembly factor that is phosphorylated by Aurora A kinase and recruited to spindle microtubules through interactions with TPX2 and importin-beta. At kinetochore fibers, DLGAP5 stabilizes microtubule attachments and facilitates KIF11 (Eg5)-mediated spindle pole separation, thereby promoting faithful chromosome segregation. The gene is transcriptionally upregulated by E2F1 and the FOXM1 transcription factor network, positioning it within a cell cycle?Cregulated circuit that also includes PLK1, Cyclin B1-CDK1, and the APC/C-CDC20 complex. DLGAP5 further interacts with DLG1/SAP97, and its activity indirectly stabilizes Cyclin B1. Knockout of DLGAP5 removes this critical stabilizing function, leading to catastrophic spindle defects, sustained mitotic arrest, chromosomal instability, and subsequent apoptosis.
In the Jurkat T lymphocyte context, loss of DLGAP5 is particularly instructive because it couples mitotic dysfunction with the inherent signaling and survival pathways of leukemic cells. Disruption of spindle integrity in these cells can reveal dependencies on mitotic checkpoint signaling, offering a model for chromosomal instability syndromes and acute T cell leukemia progression. The polyclonal nature of the knockout population allows researchers to assess heterogeneous responses and identify emergent phenotypes that may be masked in clonal isolates, making it a valuable tool for studying the interplay between oncogenic transformation and mitotic fidelity.
This product supports a wide array of experimental workflows. Researchers can measure DLGAP5, FOXM1, and E2F1 transcript levels by quantitative RT-PCR, verify protein expression and phosphorylation (e.g., phospho-Aurora A Thr288, Cyclin B1) by immunoblotting, and evaluate spindle morphology via immunofluorescence staining for ??-tubulin and pericentrin. Functional assays include flow cytometry for DNA content and mitotic index (phospho-Histone H3 Ser10), time-lapse microscopy to track mitotic progression, and annexin V apoptosis assays. Co-immunoprecipitation experiments can probe interactions with Aurora A or TPX2, while chromosome spread analysis reveals aneuploidy. These applications facilitate dissection of mitotic spindle assembly mechanisms, exploration of chromosomal instability in cancer, testing of anti-mitotic drug sensitivity, elucidation of synthetic lethality in DLGAP5-deficient cells, and investigation of DLGAP5??s role in T cell leukemia proliferation. For additional information or to discuss custom projects, please contact Ascent Research.
