The CD46 Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma cell line, engineered to disrupt the CD46 gene. This polyclonal model offers a heterogeneous genetic background, recapitulating population-level variation in gene disruption and enabling robust functional assays. The CRISPR/Cas9 system introduces targeted gene modifications, resulting in loss of CD46 expression without isolating single-cell clones. As a polyclonal population, these cells are well-suited for studying collective cellular responses to gene knockout under physiologically relevant conditions, providing a versatile platform for immunological and virological investigations.
The host cell line, Huh-7, is an established human hepatocellular carcinoma model derived from a well-differentiated liver tumor. Huh-7 cells retain key hepatic epithelial characteristics, including albumin secretion and cytochrome P450 activity, making them a widely adopted system for hepatocyte biology, drug metabolism, and liver cancer research. These cells are permissive to various pathogens, notably measles virus and adenovirus, which exploit CD46 as a cellular receptor. The Huh-7 background thus provides a clinically relevant context for dissecting the interplay between complement regulation, viral entry, and tumor immune evasion in a hepatic environment.
CD46, also known as membrane cofactor protein (MCP), is a type I transmembrane glycoprotein that protects host cells from complement-mediated lysis by serving as a cofactor for Factor I-mediated proteolytic cleavage of C3b and C4b. CD46 is a receptor for measles virus through interaction with the viral hemagglutinin (H) glycoprotein and for adenovirus via binding to the fiber knob protein. CD46 expression is upregulated by inflammatory cytokines such as tumor necrosis factor-alpha (TNF-??) and interleukin-1 beta (IL-1??), and upon viral infection. The protein interacts with C3b, C4b, measles H glycoprotein, adenovirus fiber, SPAK, and CD9, and participates in signaling pathways involving extracellular signal-regulated kinase (ERK) activation and interleukin-10 (IL-10) secretion. Downstream, CD46 modulates T-cell differentiation and limits complement deposition, thereby integrating innate and adaptive immune responses. In the complement cascade, CD46 cooperates with Complement Factor I and components C3 and C4 to degrade opsonins and prevent membrane attack complex formation.
In the context of Huh-7 hepatocellular carcinoma cells, disruption of CD46 removes a critical safeguard against complement-dependent cytotoxicity, rendering the knockout cells highly susceptible to autologous complement attack. This vulnerability recapitulates features of atypical hemolytic uremic syndrome, where CD46 deficiency leads to uncontrolled complement activation. Moreover, CD46 ablation blocks the entry of measles virus and adenovirus, providing a clean genetic system to isolate viral infection mechanisms and host dependency factors. The knockout also mirrors immune evasion strategies in hepatocellular carcinoma, where CD46 overexpression often protects malignant cells from complement surveillance and promotes an immunosuppressive microenvironment via IL-10 induction. Consequently, this model enables studies on how CD46 loss reshapes tumor?Cimmune interactions and influences hepatocyte survival under inflammatory stress.
The CD46 Knockout Huh-7 Polyclonal Cells are tailored for advanced biomedical research applications, including complement regulation studies, viral entry and infection modeling, cancer immune evasion investigation, and drug screening for complement-mediated therapies. Typical assays using this model include Western blotting and flow cytometry for confirming CD46 depletion, complement cytotoxicity assays to quantify lytic susceptibility, measles virus infection assays to evaluate viral receptor dependence, and RT-qPCR for assessing downstream gene expression changes. This product is also amenable to high-throughput screening platforms aimed at identifying modulators of complement activity or viral inhibition in hepatocellular carcinoma contexts. For technical inquiries, ordering information, and additional support, please contact Ascent Research.
