CD247 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the CD247 gene within the HAP1 cell line. This product provides a loss-of-function model for investigating the role of the CD3?? protein in T-cell receptor (TCR) signaling and immune regulation. The polyclonal format ensures a heterogeneous mixture of edited alleles, enabling robust functional studies without clonal artifacts. By targeting CD247, these cells facilitate the examination of TCR complex assembly, signal initiation, and downstream effector cascades in a convenient and genetically stable system.
The HAP1 host cell line is a near-haploid, adherent model derived from KBM-7 chronic myeloid leukemia. Its haploid karyotype reduces genetic redundancy, allowing unambiguous interpretation of knockout phenotypes and reducing compensatory effects often observed in diploid cells. HAP1 cells are widely used for functional genomics, drug screening, and mechanistic studies due to their rapid growth and amenability to genetic manipulation. The male origin and myeloid background provide a consistent context for dissecting gene function in signal transduction and disease research.
CD247 encodes CD3??, a critical TCR subunit that pairs with CD3??, CD3??, CD3??, and TCR????. Upon TCR engagement by peptide-MHC, Src kinases Lck and Fyn phosphorylate ITAMs in CD3??, recruiting ZAP-70. ZAP-70 phosphorylates LAT and SLP-76, activating PLC??1, MAP kinases (ERK, JNK, p38), NFAT, and NF-??B. Thus, CD3?? is a master regulator of T-cell activation, proliferation, and effector function. Its disruption abolishes TCR-proximal signaling, providing a clean background for pathway dissection.
In the HAP1 near-haploid system, CD247 knockout creates a simplified platform for studying TCR signaling. Although HAP1 cells are not of T-cell origin and lack endogenous TCR expression, they can be engineered with individual TCR components for reconstitution experiments. This allows assessment of CD3?ơ?s specific contribution to receptor assembly and signal transduction without interference from lymphoid-specific factors. The model recapitulates features of CD3?? deficiency, a severe primary immunodeficiency, and supports research into T-cell lymphomas and autoimmune dysregulation.
Typical applications include flow cytometry for CD3 expression, western blotting for CD3?? and phospho-ZAP-70, IL-2 ELISA, NFAT reporter assays, co-immunoprecipitation, and RNA-seq. These cells accelerate TCR signaling studies, immunodeficiency research, CAR-T cell development, and autoimmunity investigations. For more information, contact Ascent Research.
