PLAUR Knockout HAP1 Cell Line

The choice depends on whether you are studying PLAUR (urokinase plasminogen activator receptor, uPAR/CD87)'s role in pericellular plasmin generation and cancer invasion or its emerging functions as a signaling receptor. The Knockout line is the standard tool for asking whether uPAR is required for these processes — uPAR is a GPI-anchored receptor that binds uPA (urokinase) and concentrates plasmin-generating proteolytic activity at the cell surface, with established roles in cancer invasion, metastasis, and inflammation. Overexpression is useful for studying uPAR in cancer contexts. For cancer biology research, the EDITGENE PLAUR Knockout in HAP1 is a clean genetic background for uPAR functional studies. Rescue with wild-type or signaling-deficient uPAR enables structure-function studies. The knockout is a critical specificity control for uPAR-targeted therapeutics including uPAR-CAR T cells (in clinical development for senescent cell clearance and oncology) and small-molecule uPA-uPAR interaction inhibitors. uPAR is also a target of clinical antibody MNRP1685A and similar agents.

Primary applications: • Pericellular plasmin generation: cell surface uPA-uPAR-plasminogen activation assays to quantify uPAR-dependent plasmin generation. • Cancer invasion: 3D invasion assays and pericellular matrix degradation given uPAR's role in tumor invasion. • Senescent cell biology: uPAR is upregulated on senescent cells — uPAR-CAR T cells specifically target senescent cells in pre-clinical models. • uPAR-targeted therapeutic specificity: critical genetic control for uPAR-CAR T cells, MNRP1685A antibody, and small-molecule uPA-uPAR inhibitors. EDITGENE recommends this model for researchers investigating urokinase plasminogen activator receptor biology, cancer invasion mechanisms, and senotherapy or uPAR-targeted drug development.

Yes. uPAR rescue experiments require attention to GPI-anchored protein topology: • Construct design: use a codon-modified PLAUR sequence with a small N-terminal tag (FLAG, HA) — uPAR is a GPI-anchored protein with C-terminal GPI signal that is cleaved during processing, so C-terminal tags will be lost. • Surface localization validation: confirm GPI-anchored plasma membrane localization by cell surface staining and PI-PLC sensitivity before functional assays. • Domain-deletion rescue: D1-only (uPA-binding) versus full-length uPAR rescue enables dissection of uPA-binding from signaling functions. • Functional readout: rescue should restore uPA binding (flow cytometry with uPA fluorescent probe), uPAR-CAR T cell targeting, and pericellular plasmin generation. HAP1-specific considerations: • Diploidization: HAP1 cells gradually diploidize during extended culture — confirm ploidy by flow cytometry at the time of phenotypic assay. • Integration site sensitivity: position effects on transgene expression are more pronounced in near-haploid backgrounds; generating multiple independent rescue clones is strongly recommended. • Transduction efficiency: HAP1 transduces with lentivirus at moderate efficiency — increase MOI compared to standard immortalized lines.