CD3E Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated from the DLD-1 human colorectal adenocarcinoma cell line, with targeted disruption of the CD3E gene. This polyclonal pool consists of a heterogeneous mixture of cells carrying diverse mutations at the CD3E locus, providing a versatile loss-of-function model for investigating CD3E function outside its native T-cell environment and avoiding clonal selection biases.
The host DLD-1 cell line originates from a Duke??s type C colorectal adenocarcinoma in a male patient and is widely used in cancer biology studies. DLD-1 cells are epithelial in origin and exhibit classical colorectal cancer features, including dysregulated WNT signaling and chromosomal instability. Their non-hematopoietic nature enables unique exploration of CD3E in an epithelial context, where its expression is typically absent.
CD3E encodes the CD3 epsilon subunit of the T-cell receptor (TCR)?CCD3 complex, critical for TCR signaling in T lymphocytes. CD3E associates with TCR ??/?? chains, CD3??, CD3??, and CD247 to transduce antigen recognition signals. Upon TCR engagement by peptide?CMHC, Src family kinases LCK and FYN phosphorylate CD3E ITAMs, recruiting ZAP70. This initiates a cascade through LAT and SLP-76, activating MAPK, NF-??B, and NFAT pathways that promote IL-2 transcription and T-cell activation.
In DLD-1 cells, CD3E knockout serves as an unconventional model for studying off-target effects and antibody specificity, because CD3E expression is typically restricted to T cells. This polyclonal knockout pool is an ideal negative control for immunodetection methods like western blotting and flow cytometry, ensuring that observed signals are T-cell-specific. Additionally, the model supports cancer immunotherapy research by helping assess cross-reactivity of CD3-targeted therapeutics with colorectal carcinoma.
Typical applications include western blotting and RT-qPCR to confirm CD3E ablation, flow cytometry for surface or intracellular staining, immunofluorescence for localization studies, and cell viability assays to test epithelial cell dependencies. The polyclonal nature suits population-scale analyses and antibody validation. For further technical details or ordering information, please contact Ascent Research.
