The CCDC50 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the Jurkat human T lymphocyte line, featuring targeted disruption of the CCDC50 gene. This product provides a loss-of-function model for investigating the regulatory roles of CCDC50 in autophagy, NF-??B, and Wnt signaling cascades. The polyclonal format represents a heterogeneous pool of cells with diverse editing outcomes, enabling functional studies that are not influenced by clonal selection artifacts often encountered in monoclonal knockout lines. As a ready-to-use knockout tool, these cells are well-suited for transient and stable expression experiments and high-content screening assays.
Jurkat cells are an immortalized human T-cell line derived from a T-ALL patient, widely employed as a model for T-cell receptor signaling, apoptosis, and oncogenesis. These suspension cells retain key T-lymphocyte markers and are highly permissive to genetic manipulation. Their well-characterized signaling networks, including NF-??B and Wnt pathways, make them ideal for mechanistic studies of immune function and leukemia. The Jurkat background offers robust reproducibility and compatibility with diverse biochemical and imaging assays.
CCDC50 functions as a selective autophagy receptor that bridges ubiquitinated signaling proteins to autophagosomes for lysosomal degradation, thereby negatively regulating NF-??B and Wnt pathways. It recognizes ubiquitinated NEMO and Dvl2 via ubiquitin-binding domains and interacts with LC3/GABARAP proteins through its LIR motif, often cooperating with p62/SQSTM1. This targets the cargo-CCDC50 complexes to autophagosomes, leading to degradation of NEMO and Dvl2 and attenuation of downstream signaling. Cellular stress and ubiquitination signals trigger CCDC50-mediated recruitment; its disruption causes accumulation of these signaling mediators and sustained pathway activity.
In Jurkat T cells, CCDC50 knockout relieves negative regulation of NF-??B and Wnt signaling, likely enhancing pathway activation upon stimulation. As NF-??B controls T-cell activation and survival and Wnt contributes to leukemogenesis, this model enables study of how autophagy-dependent signal termination impacts immune cell behavior and leukemia progression. It is particularly valuable for exploring autophagy-inflammation crosstalk in T-ALL and for testing pathway-targeted therapeutics. The polyclonal population also permits assessment of response heterogeneity without clonal bias.
These polyclonal knockout cells are ideal for Western blotting of LC3 and p62 to monitor autophagy flux, co-immunoprecipitation of CCDC50 with NEMO, Dvl2, or LC3, and NF-??B luciferase reporter assays. Immunofluorescence microscopy visualizes autophagosome formation, and flow cytometry enables apoptosis and surface marker analysis. Applications span autophagy research, cancer biology, immune signal transduction, and drug discovery for NF-??B- and Wnt-driven diseases. Researchers may contact Ascent Research for further details.
