The CD47 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population in which the CD47 gene has been disrupted. This heterogeneous pool of HEK293T cells carries a variety of loss-of-function alleles, enabling investigation of CD47 function without clonal selection bias. By eliminating CD47 protein expression, the model permits systematic analysis of phagocytosis inhibition, cell adhesion, and related signaling pathways in a robust cellular context.
The host cell line HEK293T is an immortalized human embryonic kidney epithelial line that stably expresses the SV40 large T antigen, enhancing plasmid replication and protein production. Originally subcloned from HEK293 cells, HEK293T is widely used for viral packaging, recombinant protein expression, and gene editing studies due to its high transfection efficiency and consistent growth characteristics. This well-characterized background ensures reproducible knockout generation and reliable assay performance.
CD47 is a cell surface glycoprotein that functions as a key immune checkpoint by engaging SIRP?? on macrophages to deliver a ??don??t eat me?? signal that suppresses phagocytosis. Beyond immune evasion, CD47 regulates cell adhesion and migration through direct interactions with integrins ??v??3 and ??IIb??3, as well as the matricellular protein thrombospondin-1; it also partners with VEGFR2 to influence angiogenic signaling. Transcription factors such as MYC, HIF-1??, NF-??B, and TNF-?? drive CD47 expression. Upon ligand binding, CD47 recruits the tyrosine phosphatases SHP-1 and SHP-2, which act downstream of SIRP?? to modulate effectors including myosin-II, Rac1, and focal adhesion kinase (FAK), ultimately interfacing with PI3K/AKT and Rho GTPase pathways to control cytoskeletal dynamics and gene expression.
In the HEK293T background, CD47 knockout removes the dominant anti-phagocytic signal, making the cells particularly useful for isolating CD47??s intrinsic contributions to integrin-mediated adhesion and thrombospondin-1 signaling without the overlay of macrophage-derived inhibition. The lack of endogenous immune effector functions allows clear dissection of CD47-dependent cytoskeletal changes and signaling pathways. Moreover, the high transfectability of HEK293T facilitates rescue experiments with wild-type or mutant CD47 constructs for structure-function studies. When co-cultured with macrophages, these CD47-null cells serve as targets in quantitative phagocytosis assays to test therapeutic antibodies blocking the CD47-SIRP?? axis.
This product supports a broad range of applications in immuno-oncology, cell biology, and drug discovery, including phagocytosis checkpoint research, cancer immunotherapy modeling, antibody drug targeting, and analysis of cell adhesion and migration. Standard experimental workflows such as Western blotting, flow cytometry, RT-qPCR, immunofluorescence, co-immunoprecipitation, SIRP?? binding assays, and phagocytosis co-cultures are directly applicable. The polyclonal nature also benefits CRISPR validation and high-content screening studies. For technical inquiries or custom solutions, please contact Ascent Research.
