The CD3E Knockout 769-P Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human CD3E gene in the 769-P clear cell renal cell carcinoma epithelial cell line. This product is supplied as a heterogeneous pool of edited cells, each carrying targeted disruption of CD3E, and serves as a versatile tool for loss-of-function studies, antibody validation, and negative control applications in renal cancer research.
The 769-P host cell line was originally derived from a clear cell renal adenocarcinoma and is widely employed as an epithelial cell model for kidney cancer. These cells exhibit characteristics typical of clear cell renal cell carcinoma, including epithelial morphology and relevant oncogenic alterations, making them a robust platform for investigating renal tumor biology and therapeutic responses. The absence of endogenous CD3E expression in 769-P cells ensures that the knockout model provides a clean genetic background for experimental manipulation.
CD3E encodes the CD3 epsilon chain, an essential subunit of the T-cell receptor (TCR)?CCD3 complex. Upon TCR engagement by peptide-bound MHC molecules, the CD3 epsilon chain, together with CD3D, CD3G, and CD247, transmits activation signals through immunoreceptor tyrosine-based activation motifs (ITAMs). Src family kinases Lck and Fyn phosphorylate these ITAMs, leading to the recruitment and activation of ZAP70, which in turn phosphorylates LAT and SLP-76. This initiates a signaling cascade involving PLC??1, ITK, VAV1, RASGRP1, and culminates in the activation of transcription factors including NFAT, AP-1, and NF-??B, driving T-cell activation, differentiation, and effector functions.
In the context of 769-P renal epithelial cells, CD3E is not normally expressed, making this knockout model particularly useful for investigating ectopically expressed or reintroduced CD3E function. It allows researchers to distinguish CD3E-dependent effects in a non-hematopoietic background, providing a controlled system for studying non-canonical roles of CD3E beyond T-cell signaling, such as potential involvement in epithelial cell adhesion or migration. This model also serves as an ideal negative control for T-cell-specific experiments, ensuring that observed effects are due to CD3E in immune cells rather than off-target interactions in epithelial cells.
Typical research applications include use as a negative control for CD3E-targeted therapies, off-target screening of small molecules or antibodies, and validation of CD3E-specific detection reagents. The polyclonal knockout pool is suitable for assays such as western blotting, RT-qPCR, Sanger sequencing to confirm gene disruption, immunofluorescence, and flow cytometry. When CD3E is reintroduced, calcium flux and T-cell activation assays can be performed in co-culture systems. For further information or to request a quote, please contact Ascent Research.
