The DNAJC13 Knockout Jurkat Polyclonal Cells comprise a heterogeneous population of Jurkat T lymphoblasts in which the DNAJC13 gene has been disrupted by CRISPR/Cas9-mediated gene editing. This product provides a polyclonal loss-of-function model system that enables the investigation of DNAJC13-dependent cellular processes without the use of clonal isolation. The polyclonal format retains genetic diversity reflective of the original editing event, making it suitable for pooled functional assays and preliminary pathway analyses.
The Jurkat cell line is an immortalized human T lymphocyte model derived from an acute T cell leukemia patient. These cells are widely employed in immunology and cancer research to study T cell receptor signaling, cytokine production, and canonical Wnt pathway dynamics. Their robust growth characteristics and genetic tractability make them an ideal host for CRISPR-based gene disruption, allowing direct interrogation of genes implicated in immune cell function and oncogenic transformation.
DNAJC13 encodes a J-domain cochaperone that is essential for endosomal protein sorting and intracellular trafficking. Mechanistically, DNAJC13 associates with the WASH complex to facilitate retromer-mediated retrieval of cargoes, including the Wnt receptor Frizzled, from endosomes to the plasma membrane. By regulating the surface availability of Frizzled, DNAJC13 controls the activation of the canonical Wnt/??-catenin cascade. Upstream regulators such as WNT3A and RAB4A influence this process, while downstream, DNAJC13 ultimately affects ??-catenin stabilization and TCF/LEF-dependent transcription. Key interacting partners include FAM21, SNX-BAR proteins, and the retromer subcomplex, placing DNAJC13 at a critical junction between endosomal sorting and Wnt signal transduction.
In the Jurkat cellular context, DNAJC13 disruption is expected to alter Frizzled receptor trafficking, thereby impacting Wnt/??-catenin signaling, a pathway integral to T cell development, proliferation, and leukemogenesis. Aberrant Wnt signaling is a hallmark of several cancers, including T cell acute lymphoblastic leukemia. Moreover, genetic variants of DNAJC13 have been linked to Parkinson??s disease, suggesting broader roles in proteostasis and neurodegeneration. Thus, this polyclonal knockout pool provides a relevant model to dissect the pathway-specific and disease-related functions of DNAJC13 in a human T lymphoblast background.
Researchers can employ this product for a variety of experimental applications, including mechanistic studies of endosomal trafficking, Wnt pathway modulation, and disease modeling. Commonly used assays with these cells include Western blotting to assess protein expression changes, immunofluorescence microscopy to visualize subcellular localization, co-immunoprecipitation to characterize protein-protein interactions with factors such as FAM21 or retromer components, and Wnt luciferase reporter assays to quantify transcriptional activity. Flow cytometry can also be used to measure surface Frizzled receptor levels, providing a direct readout of DNAJC13??s role in receptor recycling. For further details or to discuss custom gene editing projects, please contact Ascent Research.
