This product provides a CRISPR/Cas9-edited polyclonal knockout cell population of HEK293T cells harboring targeted disruption of the CD59 gene. The polyclonal format offers a heterogeneous pool of edited cells, enabling loss-of-function studies without prior clonal isolation. This model is suitable for investigating CD59-dependent processes and complement regulation.
The HEK293T host cell line is an adenovirus 5-transformed human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen. This background supports robust protein production and high-titer virus generation, making it a versatile platform for studying gene function. HEK293T cells are widely employed in complement research due to their ease of manipulation and well-characterized responses.
CD59 encodes a glycosylphosphatidylinositol (GPI)-anchored glycoprotein that functions as a critical inhibitor of the complement membrane attack complex (MAC). Upon complement activation, CD59 binds to C8 and C9 within the assembling MAC, preventing C9 polymerization and subsequent pore formation on host cell surfaces. This protective mechanism is regulated upstream by immune stimuli such as complement activation products and inflammatory cytokines like TNF-??. By directly interacting with C8 and C9, CD59 effectively blocks terminal complement-mediated lysis. The protein operates within the complement cascade, downstream of C5b, C6, and C7 recruitment, and is essential for preserving host cell integrity against autologous complement attack.
In the HEK293T background, disruption of CD59 eliminates the cell??s intrinsic resistance to complement-dependent cytotoxicity, creating a sensitized model for studying MAC-mediated lysis. This deficiency is particularly valuable for dissecting complement regulatory networks and for modeling diseases such as paroxysmal nocturnal hemoglobinuria (PNH), where CD59 deficiency leads to erythrocyte hemolysis. Moreover, the transformed nature of HEK293T cells permits co-expression studies and high-throughput screening of complement-modulating agents, enabling detailed examination of immune evasion strategies employed by cancer cells.
Researchers can utilize these polyclonal knockout cells in complement-mediated lysis assays to quantify sensitivity to serum challenge, in flow cytometry to confirm loss of surface CD59 expression, and in immunofluorescence or western blotting to detect MAC components such as C5b-9. The model supports investigations into complement regulation, PNH pathophysiology, and tumor cell resistance to complement attack. Additional applications include screening for inhibitors of MAC assembly and exploring signaling interactions between complement and inflammatory cytokines. For further information or technical support regarding this knockout cell model, please contact Ascent Research.
