SLC31A2 Knockout HCT116 Cell Line

The choice depends on whether you are studying SLC31A2 (CTR2)'s role as an intracellular low-affinity copper transporter or its emerging functions in lysosomal copper handling. The Knockout line is appropriate for asking whether CTR2 is required for these activities — CTR2 is distinct from CTR1 (SLC31A1, the high-affinity plasma membrane copper importer) and functions primarily at endosomes/lysosomes. Overexpression is useful for studying CTR2 in heterologous expression systems or for assessing platinum chemotherapy resistance mechanisms. For copper biology research, the EDITGENE SLC31A2 Knockout in HCT 116 enables study of intracellular copper trafficking distinct from CTR1-mediated uptake. CTR2 is implicated in resistance to platinum-based chemotherapy (cisplatin, carboplatin) through reduced drug accumulation. Rescue with wild-type or transport-deficient CTR2 is the standard specificity control.

Primary applications: • Intracellular copper trafficking: subcellular copper distribution analysis using copper-sensitive fluorescent probes or ICP-MS-based compartment analysis. • Platinum chemotherapy sensitivity: dose-response analysis for cisplatin, carboplatin, and oxaliplatin in SLC31A2-null versus rescued cells. • Endosomal/lysosomal copper handling: imaging-based analysis of copper-dependent processes in intracellular compartments. • CTR1 paralog studies: SLC31A1 expression analysis to assess high-affinity versus low-affinity copper transporter functions. EDITGENE recommends this model for researchers investigating intracellular copper biology, platinum chemotherapy resistance mechanisms, and copper-dependent cellular processes.

Yes. CTR2 rescue experiments require attention to intracellular targeting: • Construct design: use a codon-modified SLC31A2 sequence with a small C-terminal tag (FLAG, HA). CTR2 has 3 transmembrane domains and intracellular trafficking determinants. • Endosomal/lysosomal localization validation: confirm intracellular localization by LAMP1 or Rab9 co-staining — CTR2 functions primarily at endosomes/lysosomes. • Transport-deficient rescue: methionine-rich motif mutations enable structure-function studies of copper coordination. • Functional readout: rescue should restore intracellular copper handling and platinum chemotherapy sensitivity profiles. HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation.