The CCDC50 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated from the human K-562 chronic myelogenous leukemia lymphoblast line. These cells feature targeted disruption of the CCDC50 gene, which encodes an adaptor protein involved in endosomal sorting and signaling. The polyclonal format captures a range of editing outcomes, providing a robust loss-of-function model without clonal artifacts. Suitable for suspension culture, they are ideal for biochemical and imaging assays.
The parental K-562 line, originally from a pleural effusion of a CML patient in blast crisis, is BCR-ABL1 positive and grows as a lymphoblast suspension culture. It is a classic model for studying hematopoietic differentiation, including erythroid and myeloid lineage commitment, and is susceptible to natural killer (NK) cell cytotoxicity. The CCDC50 knockout derivative retains these features, enabling focused investigation of gene function in a leukemic background.
CCDC50 is an adapter protein that mediates ubiquitin-dependent sorting of EGFR, promoting lysosomal degradation by interacting with EGFR, c-Cbl, and ESCRT components STAM and Hrs. It also modulates NF-??B signaling through ubiquitinated intermediates and participates in selective autophagy via binding to LC3/GABARAP and p62/SQSTM1. Thus, CCDC50 links growth factor receptor trafficking to inflammatory signaling and autophagic clearance, positioning it at the intersection of multiple pathways including EGFR/MAPK, NF-??B, and autophagy.
In K-562 cells, BCR-ABL1 oncogenic signaling drives proliferation and intersects with CCDC50-regulated pathways. Disruption of CCDC50 allows dissection of how EGFR trafficking and NF-??B activity influence leukemic phenotype, including differentiation and response to cytokines. The model is also useful for exploring whether CCDC50 impacts hematopoietic lineage commitment, given K-562??s bipotential nature. Moreover, the cells?? NK susceptibility offers a system to examine if CCDC50 affects immune recognition.
Researchers can employ these cells in Western blotting to monitor EGFR degradation, phospho-ERK, and I??B??; co-immunoprecipitation to confirm EGFR?CCCDC50 binding; and immunofluorescence to assess EGFR?Clysosome colocalization. Autophagy flux assays using LC3 turnover, flow cytometry for surface EGFR, and apoptosis/proliferation assays further characterize CCDC50 function. The polyclonal population is well suited for screening chemical probes or genetic rescue experiments. For further details, contact Ascent Research.
