The CD59 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting CD59 in the 786-O clear cell renal cell carcinoma line. This loss-of-function model disrupts the gene encoding the GPI-anchored inhibitor of the complement membrane attack complex (MAC). The polyclonal pool comprises a heterogeneous mixture of cells with gene disruptions, avoiding clonal selection bias and enabling bulk cellular analyses of complement resistance mechanisms.
The 786-O host cell line originates from a VHL-mutant clear cell renal cell carcinoma and displays adherent epithelial morphology. Loss of VHL leads to constitutive HIF stabilization, driving pseudohypoxic metabolic reprogramming and altering expression of immune modulatory factors. This genetic context makes it a pertinent system for studying tumor immune evasion and complement regulatory pathways.
CD59 obstructs MAC-mediated lysis by binding directly to C8 and C9, preventing C9 polymerization and pore formation. Transcription of CD59 is controlled by IFN-??, TNF-??, IL-1??, LPS, and hypoxia, linking its expression to inflammatory and metabolic cues. Downstream, it inhibits the C5b-9 complex and suppresses complement-dependent cytotoxicity (CDC). CD59 also interacts with CD2, and functions alongside the membrane-bound complement regulators DAF (CD55) and MCP (CD46) to safeguard host cells from autologous complement attack.
In VHL-deficient 786-O cells, CD59 knockout eliminates a principal complement defense mechanism, sensitizing tumor cells to immune effector functions. Upregulation of CD59 by hypoxia??a prominent feature of clear cell carcinoma??suggests that this polyclonal knockout system allows dissection of complement resistance pathways decoupled from HIF-dependent regulation. The model thus provides a robust platform to explore how renal cancer cells subvert complement surveillance and to test strategies for restoring CDC sensitivity.
This polyclonal knockout pool supports diverse applications including complement evasion studies, immunotherapy resistance modeling, and tumor microenvironment analysis. It is compatible with complement-dependent cytotoxicity assays, complement deposition assays, flow cytometry, immunofluorescence, and Western blotting to verify CD59 disruption and functional outcomes. These tools facilitate comprehensive investigation of complement regulatory circuits in renal cancer. For additional product information or technical support, please contact Ascent Research.
