MMP16 Knockout HAP1 Cell Line

The choice depends on whether you are studying MMP16 (MT3-MMP)'s role as a transmembrane membrane-type matrix metalloproteinase or its functions in tissue remodeling and pro-MMP-2 activation. The Knockout line is the standard tool for asking whether MMP16 is required for these processes — MMP16 is a type I transmembrane MT-MMP that activates pro-MMP-2 at the cell surface and degrades various ECM substrates (collagen III, fibronectin, vitronectin). Overexpression is useful for studying MMP16 in heterologous expression contexts. For matrix metalloproteinase research, the EDITGENE MMP16 Knockout in HAP1 enables study of MT3-MMP biology. Other MT-MMP (MMP14, MMP15) expression analysis is essential given functional overlap in pro-MMP-2 activation. Rescue with wild-type or catalytically-dead MMP16 is the standard specificity control. This product complements parallel MMP15 and MMP25 Knockouts in HAP1 for MT-MMP family functional dissection.

Primary applications: • pro-MMP-2 activation: zymography or activity assays to characterize MMP16-dependent pro-MMP-2 cleavage to active MMP-2. • ECM degradation: collagen III, fibronectin, vitronectin substrate degradation assays. • MT-MMP family comparison: parallel analysis with MMP15 and MMP25 Knockouts in HAP1 for paralog-specific MT-MMP characterization. • Cell invasion: 3D invasion assays in ECM-rich matrices given MT-MMP's role in pericellular proteolysis. EDITGENE recommends this model for researchers investigating MT-MMP family biology and pericellular ECM remodeling.

Yes. MMP16 rescue experiments require attention to transmembrane topology: • Construct design: use a codon-modified MMP16 sequence with a small intracellular C-terminal tag (FLAG, HA). MMP16 has N-terminal signal/prodomain, extracellular catalytic domain, hinge, hemopexin domain, single transmembrane span, and short cytoplasmic tail — preserve all elements. • Catalytically-dead rescue: HEXXH motif glutamate mutation (E to A) abolishes metalloprotease activity. • Prodomain processing: active MMP16 is generated by furin processing of the prodomain — confirm mature MMP16 form after rescue. • Functional readout: rescue should restore pro-MMP-2 activation (zymography) and ECM substrate degradation. 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.