The CD47 Knockout 786-O Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human 786-O renal epithelial carcinoma cell line, designed to disrupt expression of the immune checkpoint gene CD47. This genetically heterogeneous pool provides a loss-of-function model for investigating CD47-dependent signaling, phagocytosis regulation, and integrin-mediated adhesion without the constraints of clonal selection. As a polyclonal population, these cells retain the biological variability inherent to CRISPR/Cas9-mediated gene disruption, offering a robust and reproducible system for functional studies.
The 786-O parental line originates from a primary clear cell renal cell carcinoma (ccRCC) and harbors a mutant VHL tumor suppressor, leading to constitutive activation of hypoxia-inducible factor HIF-1??. These adherent cells serve as a well-characterized model for ccRCC, a malignancy characterized by immune evasion and aberrant integrin signaling. The VHL-deficient background recapitulates key oncogenic drivers, including upregulation of HIF-1?? target genes, making 786-O an ideal host for dissecting CD47??s contribution to tumor biology in a therapeutically relevant context.
CD47 encodes an integrin-associated protein that functions as a critical immune checkpoint by binding to signal regulatory protein alpha (SIRP??) on macrophages. This interaction triggers SHP-1 and SHP-2 phosphatase-mediated dephosphorylation, inhibiting myosin-IIA accumulation at the phagocytic synapse and thereby delivering a ??don??t eat me?? signal. CD47 additionally interacts with integrins such as ??v??3 and ??2??1, thrombospondin-1, and VEGFR2, linking it to cell adhesion, migration, and integrin signaling. Upstream, CD47 expression is regulated by HIF-1??, NF-??B, and MYC, while downstream, CD47-SIRP?? engagement suppresses the PI3K/Akt pathway and Rho GTPase activity, influencing cytoskeletal dynamics and cell survival.
In 786-O cells, disruption of CD47 by CRISPR/Cas9 abrogates the protective anti-phagocytic signal, potentially enhancing susceptibility to macrophage-mediated clearance and altering immune surveillance. This knockout model also perturbs CD47??s role in integrin-dependent adhesion and migration, processes often deregulated in ccRCC. Given the VHL-mutant background driving HIF-1?? overexpression, these cells enable direct investigation of CD47 regulation under hypoxic signaling, a common feature of solid tumors. Consequently, the polyclonal knockout population is invaluable for studying CD47-driven immune evasion and tumor microenvironment interactions in renal carcinoma.
Researchers can employ these CD47 knockout polyclonal cells in a diverse range of assays, including phagocytosis assays with macrophages, SIRP?? binding studies, and flow cytometry to validate CD47 loss. Western blotting and immunofluorescence further enable assessment of downstream signaling components like SHP-1, SHP-2, and PI3K/Akt pathway members. Drug screening for CD47 antagonists, migration/invasion assays, and co-culture systems with immune cells are directly applicable. For more information or to explore how this CD47 Knockout 786-O Polyclonal Cell population can advance your research, please contact Ascent Research.
