CD3E Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human tongue squamous cell carcinoma cell line CAL-27. This product features targeted disruption of the CD3E gene, encoding the epsilon subunit of the T-cell receptor CD3 complex. The polyclonal format provides a heterogeneous pool of edited cells, enabling robust loss-of-function studies without monoclonal selection bottlenecks. These cells are suitable for applications in immunology and cancer research.
The CAL-27 cell line originates from human oral squamous cell carcinoma of the tongue, representing an adherent epithelial model widely used in cancer biology and drug screening. CAL-27 cells exhibit typical epithelial morphology and retain key signaling pathways relevant to oncogenic transformation, invasion, and metastasis. Although non-hematopoietic, these cells may ectopically express immune-related genes, making them a useful platform for investigating non-canonical immune receptor functions or as a control in CRISPR off-target analyses.
CD3E encodes the CD3-epsilon chain, an essential component of the T-cell receptor (TCR) complex that forms heterodimers with CD3G and CD3D and associates with CD247 homodimer to transduce antigen recognition signals. In T lymphocytes, antigen engagement triggers phosphorylation of ITAMs by Lck kinase, recruiting and activating ZAP70 kinase. ZAP70 then phosphorylates LAT and PLCgamma1, initiating MAPK, NF-kappa B, and PI3K-Akt pathways that activate transcription factors NFAT, NF-kB, and AP-1, leading to IL-2 production. CD3E is a critical hub in T-cell activation, development, and adaptive immunity, with CD4 and CD8 coreceptors stabilizing TCR-pMHC interactions.
In the CAL-27 epithelial cancer cell line, CD3E expression is not part of the native TCR signaling machinery. Its knockout here is valuable for studying potential off-target CRISPR effects or non-canonical roles of CD3-related proteins. The polyclonal knockout pool mimics tumor genetic heterogeneity and allows assessment of gene disruption efficiency across a population. This model can serve as a negative control in T-cell functional assays or for validating antibodies and probes targeting CD3E, where residual immunoreactivity indicates nonspecific binding.
These knockout cells support diverse applications. They enable knockout validation by Western blotting, flow cytometry, and RT-qPCR to confirm CD3E loss. Functional assays such as MTT proliferation, wound healing, and transwell invasion assess potential CD3E effects on epithelial behavior. They serve as a negative control for T-cell receptor signaling studies and a system to evaluate off-target CRISPR effects. In immunotherapy, they test specificity of CD3-bispecific antibodies or CAR-T cell constructs. For further details or to request a quotation, please contact Ascent Research.
