CD2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of HEK293T cells with targeted disruption of the human CD2 gene. The polyclonal pool contains heterogeneous edited alleles, providing a robust loss-of-function model without clonal selection artifacts. CRISPR/Cas9-mediated gene disruption eliminates CD2 protein expression, enabling gene-function studies.
The HEK293T host cell line is derived from human embryonic kidney cells transformed with adenovirus 5 DNA and expresses SV40 large T antigen. These adherent epithelial cells are widely used for high-level heterologous gene expression and lentiviral production due to their high transfection efficiency and rapid growth. Their well-characterized biology ensures reliable experimental performance.
CD2 is a T-cell surface glycoprotein that mediates adhesion and costimulatory signaling. It binds CD58 (and with lower affinity CD48) on antigen-presenting cells, recruiting the Src kinases Lck and Fyn, which phosphorylate LAT, SLP-76, and PLC??1 to activate RAS?CRAF?CMEK?CERK (MAPK1/3) and PI3K?CAKT cascades. Downstream, transcription factors NFAT, AP-1, and NF-??B drive IL-2 gene expression. CD2 expression is regulated by TCR signaling, IL-2, NFAT, AP-1, and ETS transcription factors. The cytoplasmic adaptor CD2AP connects CD2 to the actin cytoskeleton.
HEK293T cells lack endogenous CD2, so the knockout ensures a null background for heterologous expression studies. Since CD2 is a T-cell?Cspecific protein, its knockout in a non-immune cell line creates an isolated system for studying CD2-dependent signals without confounding endogenous T-cell pathways. This polyclonal model is ideal for reconstitution experiments with wild-type or mutant CD2 to dissect adhesion and signaling mechanisms. It reduces background in assays such as CD58 adhesion and kinase recruitment, and the polyclonal nature minimizes clonal variability, enhancing reproducibility.
These cells support diverse applications: western blotting and RT?qPCR for CD2 depletion verification, immunocytochemistry for localization, flow cytometry for knockout confirmation, and adhesion assays with CD58-coated surfaces. They enable protein interaction studies with Lck, Fyn, CD58, and CD2AP, as well as functional complementation and luciferase reporter assays for NFAT/AP-1 activation. The cells serve as a control line for CD2 expression experiments and as a negative control to confirm assay specificity. For further details, contact Ascent Research.
