CD46 and CD55 and CD59 Knockout HEK293 Cell Line

The choice depends on whether you are studying combined complement regulation or distinguishing the contributions of the three membrane-bound complement regulators. The Triple Knockout line is uniquely valuable for asking whether CD46/CD55/CD59 are required for complement regulation — CD46 (membrane cofactor protein, MCP), CD55 (decay-accelerating factor, DAF), and CD59 (protectin) constitute the three principal membrane-bound complement regulatory proteins (CRPs); CD46 functions as a cofactor for factor I-mediated cleavage of C3b/C4b, CD55 accelerates the decay of C3/C5 convertases, and CD59 inhibits MAC (membrane attack complex) assembly by binding C8 and C9. Combined triple knockout completely eliminates membrane CRP regulation, generating maximally complement-sensitive cells. For complement biology and antibody-dependent cellular cytotoxicity (CDC) research, the EDITGENE CD46 & CD55 & CD59 Triple Knockout in HEK293 is the gold-standard genetic tool — single or double knockouts retain residual CRP activity; triple knockout completely abolishes membrane-anchored complement regulation. Single-isoform rescue (CD46 alone, CD55 alone, or CD59 alone) enables isoform-specific functional dissection — the gold-standard experimental design for the three-CRP system. The triple knockout is uniquely valuable for studying ⭐ rituximab, obinutuzumab, ofatumumab CDC mechanisms (these anti-CD20 antibodies eliminate B-cell lymphomas partially via CDC), eculizumab/ravulizumab pharmacology (anti-C5 antibodies for PNH/aHUS), xenotransplantation research (porcine CRPs are not effective against human complement), and emerging CDC-enhanced therapeutic antibody development.

Primary applications: • Complement-dependent cytotoxicity (CDC): in heterologous antibody-treated systems, CDC activity should be dramatically enhanced in the triple KO given complete loss of membrane CRP regulation. • Anti-CD20 antibody CDC mechanism: critical genetic background for studying ⭐ rituximab, obinutuzumab, ofatumumab CDC contribution to B-cell lymphoma elimination. • Single-isoform rescue: re-introduction of CD46, CD55, or CD59 alone enables systematic functional dissection — gold-standard experimental design. • Eculizumab/ravulizumab mechanism: in heterologous PNH/aHUS-relevant contexts, anti-C5 antibody mechanism studies. • Xenotransplantation research: porcine cells require human CRP rescue to evade human complement — this triple KO provides the human CRP-null platform. EDITGENE recommends this triple knockout as the gold-standard genetic tool for complement biology, CDC-enhanced antibody therapy development, and xenotransplantation research.

Yes, and rescue experiments are uniquely powerful in this triple knockout: • Single-isoform rescue: re-introduction of CD46, CD55, or CD59 alone in the triple knockout enables systematic functional dissection — gold-standard experimental design for the three membrane CRPs. • Construct design: use codon-modified sequences with small tags. CD46 (type I membrane protein) — small intracellular C-terminal tag (FLAG, HA); CD55 (GPI-anchored) — small N-terminal tag (after signal peptide; C-terminal GPI signal cleaved); CD59 (GPI-anchored) — small N-terminal tag (similar GPI processing). • Surface localization validation: confirm plasma membrane localization for each CRP before complement assays. • Pairwise rescue: combinations of two CRPs enable understanding of CRP cooperation. • Functional readout: rescue should restore CRP-specific complement protection — CD46 should rescue C3b/C4b inactivation, CD55 should rescue convertase decay, CD59 should rescue MAC inhibition. HEK293 transduces efficiently with lentivirus and supports systematic isoform-specific rescue experiments for the foundational complement regulation field.