CA9 Knockout Cell Line (HEK 293)

The choice depends on whether you are studying CA9 (carbonic anhydrase IX, CAIX)'s role as the principal hypoxia-induced carbonic anhydrase or modeling its applications as a renal cell carcinoma biomarker and therapy target. The Knockout line is the standard tool for asking whether CA9 is required for these processes — CA9 is a HIF1α-induced surface carbonic anhydrase that catalyzes CO₂ + H₂O ↔ HCO₃⁻ + H⁺ at the extracellular face, acidifying the tumor microenvironment and supporting cancer cell pH regulation under hypoxia; CA9 is uniquely restricted to gastric epithelium under normal conditions but dramatically upregulated in many hypoxic tumors. Overexpression is useful for studying CA9 gain-of-function effects. For cancer hypoxia research, the EDITGENE CA9 Knockout in HEK293 is uniquely valuable — CA9 is one of the most validated hypoxic tumor biomarkers and emerging cancer therapy targets, particularly in renal cell carcinoma (ccRCC has near-universal CA9 expression due to VHL loss-induced HIF1α stabilization). Rescue with wild-type or catalytically-dead CA9 enables structure-function studies. The knockout is a critical specificity tool for ⭐ girentuximab (anti-CA9 antibody) — recently completed Phase III for ccRCC, ⁸⁹Zr-girentuximab PET imaging (CA9-targeted ccRCC imaging), Lu-177-girentuximab radioligand therapy candidates, anti-CA9 CAR-T cells, and CA9-selective inhibitors (acetazolamide is non-selective; selective CA9 inhibitors SLC-0111 in clinical development).

Primary applications: • Hypoxia signaling: HIF1α-induced CA9 expression analysis under normoxia/hypoxia/CoCl₂ stress in CA9-null cells. • CA9-targeted therapy specificity: critical genetic control for ⭐ girentuximab (Phase III ccRCC), ⁸⁹Zr-girentuximab PET imaging, Lu-177-girentuximab radioligand therapy candidates, anti-CA9 CAR-T cells. • CA9-selective inhibitor specificity: SLC-0111 and other emerging CA9-selective inhibitors. • Cancer pH regulation: extracellular pH and intracellular pH analysis given CA9's role in pHe acidification. EDITGENE recommends this model as a critical specificity control for CA9-targeted ccRCC therapy and cancer hypoxia research.

Yes. CA9 rescue experiments require attention to membrane targeting and dual-domain architecture: • Construct design: use a codon-modified CA9 sequence with a small intracellular C-terminal tag (FLAG, HA). CA9 has N-terminal proteoglycan-like (PG) domain, central catalytic carbonic anhydrase domain, single transmembrane span, and short cytoplasmic tail — preserve membrane topology. • Surface localization validation: confirm plasma membrane CA9 by cell surface staining before girentuximab/SLC-0111 testing. • Catalytically-dead rescue: active site zinc-binding histidine mutations abolish carbonic anhydrase activity. • Functional readout: rescue should restore CA9 enzymatic activity and girentuximab binding. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.