CD274 Knockout Cell Line (Hela)

The choice depends on whether you are studying CD274 (PD-L1)'s role as the principal immune checkpoint ligand or modeling PD-L1-targeted cancer immunotherapy in imaging-friendly context. The Knockout line is the standard tool for asking whether PD-L1 is required for these processes — PD-L1 (CD274, B7-H1) is the principal ligand for PD-1 (PDCD1) on activated T cells; PD-L1-PD-1 engagement transduces inhibitory signaling through PD-1 ITSM motifs, recruiting SHP1/SHP2 to dephosphorylate TCR signaling components and dampen T cell activation; PD-L1 is constitutively expressed at low levels and dramatically upregulated by IFN-γ via JAK-STAT-IRF1 signaling; PD-L1 expression on tumor cells is a major immune evasion mechanism. Overexpression is useful for studying PD-L1 gain-of-function effects. For PD-L1 cancer immunotherapy research, the EDITGENE CD274 Knockout in HeLa is uniquely valuable — HeLa's flat morphology supports imaging-based PD-L1 trafficking studies. This product complements the parallel CD274 Knockouts in HEK293 (biochemistry) and HCT 116 (colorectal context) for cross-background validation. Rescue with wild-type or trafficking-deficient PD-L1 enables structure-function studies. The knockout is a critical specificity control for ⭐⭐⭐ all PD-L1-targeted therapies: ⭐⭐⭐ atezolizumab (Tecentriq), durvalumab (Imfinzi), avelumab (Bavencio), and the anti-PD-1 antibodies that target the PD-L1 partner — pembrolizumab (Keytruda, top-selling cancer drug), nivolumab (Opdivo), cemiplimab (Libtayo); also CMTM6-targeted approaches (CMTM6 stabilizes PD-L1).

Primary applications: • IFN-γ-induced PD-L1: IFN-γ-stimulated PD-L1 induction analysis by flow cytometry and Western blot — CD274 KO eliminates this canonical PD-L1 upregulation. • Imaging-based PD-L1 trafficking: confocal imaging of PD-L1 surface expression, internalization, and recycling given HeLa's flat morphology. • PD-L1-targeted therapy specificity: critical genetic control for atezolizumab (Tecentriq), durvalumab (Imfinzi), avelumab (Bavencio). • PD-1-PD-L1 axis dissection: in heterologous T cell co-culture systems, PD-L1-mediated T cell suppression analysis. EDITGENE recommends this HeLa-based model for imaging-based PD-L1 research; the parallel HEK293 and HCT 116 KOs (also available) are preferred for biochemistry and MSI-H cancer studies respectively.

Yes. PD-L1 rescue experiments are well-established for cancer immunotherapy research: • Construct design: use a codon-modified CD274 sequence with a small intracellular C-terminal tag (FLAG, HA). PD-L1 is a type I membrane protein with extracellular IgV and IgC2 domains (PD-1 binding), transmembrane span, and short cytoplasmic tail — preserve membrane topology. • Surface localization validation: confirm plasma membrane PD-L1 by cell surface staining (anti-PD-L1 antibody clones 28-8, SP142, 22C3, SP263) before functional assays. • PD-1-binding-deficient rescue: IgV domain mutations abolish PD-1 binding while preserving surface expression. • Functional readout: rescue should restore IFN-γ-inducible PD-L1 expression and anti-PD-L1 antibody/anti-PD-1 antibody activity in PD-1-expressing T cell co-cultures. HeLa transduces efficiently with lentivirus and supports stable rescue line generation for systematic PD-L1 imaging studies.