CTLA4 Knockout Cell Line (HEK 293)

The choice depends on whether you are studying CTLA4 (cytotoxic T-lymphocyte-associated antigen 4, CD152)'s role as the prototypical immune checkpoint or modeling CTLA-4-targeted cancer immunotherapy. The Knockout line is the standard tool for asking whether CTLA-4 is required for these processes — CTLA-4 is a transmembrane immune checkpoint receptor expressed on activated T cells and constitutively on regulatory T cells (Tregs); CTLA-4 binds CD80/CD86 on antigen-presenting cells with much higher affinity than the co-stimulatory CD28, sequestering these ligands and downregulating T cell activation; CTLA-4 also trans-endocytoses CD80/CD86 from APCs via Treg function. Overexpression is useful for studying CTLA-4 in heterologous expression contexts. For immune checkpoint research, the EDITGENE CTLA4 Knockout in HEK293 is uniquely valuable — HEK293 supports systematic structure-function studies of CTLA-4 and serves as a critical specificity control for anti-CTLA-4 antibody development. Rescue with wild-type or trafficking-deficient CTLA-4 enables structure-function studies. The knockout is a critical specificity tool for ⭐⭐ ipilimumab (Yervoy, FDA-approved 2011 — the first immune checkpoint inhibitor; melanoma, NSCLC, RCC, HCC, MSI-H CRC indications), tremelimumab (FDA-approved for HCC and NSCLC), and emerging next-generation CTLA-4-targeted strategies — anti-CTLA-4 therapy initiated the cancer immunotherapy revolution (James Allison's 2018 Nobel Prize).

Primary applications: • Anti-CTLA-4 antibody specificity: critical genetic control for ipilimumab (Yervoy) and tremelimumab — these antibodies should have no binding/effect in CTLA4-null cells. • CD80/CD86 ligand binding: CTLA-4-CD80/CD86 binding analysis given CTLA-4's higher affinity than CD28. • Trans-endocytosis: in heterologous co-culture systems, characterization of CTLA-4-mediated CD80/CD86 trans-endocytosis from antigen-presenting cells. • Next-generation checkpoint inhibitor specificity: emerging anti-CTLA-4 antibodies, bispecific antibodies, and ADC specificity validation. EDITGENE recommends this model as a critical specificity control for the foundational ipilimumab-tremelimumab class of immune checkpoint inhibitors.

Yes. CTLA-4 rescue experiments require attention to transmembrane checkpoint architecture: • Construct design: use a codon-modified CTLA4 sequence with a small intracellular C-terminal tag (FLAG, HA). CTLA-4 has extracellular IgV-like CD80/CD86-binding domain, single transmembrane span, and intracellular GYFIN motif (β-arrestin trafficking) — preserve all elements. • Surface localization validation: confirm plasma membrane localization by cell surface staining before functional assays — CTLA-4 is mostly intracellular at steady state. • Ligand-binding-deficient rescue: extracellular domain mutations abolish CD80/CD86 binding. • Trafficking-deficient rescue: GYFIN motif mutations affect endocytic trafficking. • Functional readout: rescue should restore CD80/CD86 binding and CTLA-4-mediated immune checkpoint function in appropriate immune cell co-culture systems. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation for ipilimumab/tremelimumab specificity testing.