The CD3E Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human renal clear cell adenocarcinoma line. This product provides a loss-of-function model for the CD3E gene, encoding the T-cell receptor epsilon chain, through targeted gene disruption. The polyclonal format offers a diverse pool of edited cells, suitable for robust phenotypic screening. The polyclonal nature ensures a broad representation of gene-disrupted cells, minimizing clonal bias.
The parental 786-O cell line originates from human renal proximal tubule epithelial cells and is a widely used model of clear cell renal cell carcinoma. It carries a mutant VHL gene and is PTEN-null, mirroring the genetic lesions common in this cancer type. These features make 786-O cells instrumental for studying tumorigenesis, hypoxia signaling, and drug resistance, now extended by this knockout derivative.
CD3E encodes the epsilon subunit of the TCR/CD3 complex, essential for T-cell activation. Upon antigen-MHC recognition, CD3E interacts with CD3 gamma, delta, and zeta chains and TCR alpha/beta to initiate signaling. Lck kinase phosphorylates CD3E ITAMs, recruiting ZAP-70, which then activates LAT and PLC-gamma1. This leads to calcium mobilization and activation of NFAT, NF-kB, and AP-1 transcription factors, driving cytokine expression such as IL-2. Transcription factors TCF-1 and GATA3 regulate CD3E upstream.
Although primarily expressed in T lymphocytes, ectopic CD3E expression may occur in certain cancer cells. In the 786-O renal carcinoma background, knockout of CD3E permits investigation of its potential non-canonical functions independent of a complete TCR complex. The intersection of CD3E-mediated calcium and MAPK pathways with VHL- and PTEN-dependent signaling may uncover novel roles in tumor biology.
Researchers can employ this model in T-cell receptor signaling studies, immunodeficiency modeling, and T-cell activation assays, especially to explore the significance of aberrant CD3E expression in renal carcinoma. Typical validation includes western blotting, RT-qPCR, flow cytometry, and genomic sequencing; functional assays such as proliferation, migration, and invasion experiments reveal consequences of CD3E loss. For further information, please contact Ascent Research.
