SLC38A5 Knockout Cell Line (HCT 116)

The choice depends on whether you are studying SLC38A5 (SNAT5)'s role in glutamine transport or its emerging functions in cancer metabolism and the malate-aspartate shuttle. The Knockout line is the standard tool for asking whether SNAT5 is required for sodium-coupled glutamine and other neutral amino acid uptake — SNAT5 (System N) is distinct from System A (SNAT1/2/4) transporters. Overexpression is useful for studying SNAT5's role in cancer cell glutamine addiction. For SNAT5 research, the EDITGENE Knockout in HCT 116 is relevant for cancer metabolism studies — colorectal cancer cells are often glutamine-dependent, and SNAT5 contributes to glutamine acquisition. SNAT3 (SLC38A3) paralog expression should be assessed given overlap in System N function. Rescue with wild-type or transport-deficient SNAT5 enables structure-function studies and substrate specificity characterization.

Primary applications: • Glutamine uptake: ³H-glutamine uptake assays to quantify SNAT5-dependent glutamine transport — System N specifically transports glutamine, asparagine, and histidine. • Cancer metabolism: glutamine-dependent proliferation and glutaminolysis assays relevant to colorectal cancer metabolism. • SNAT family studies: SNAT3 (SLC38A3) expression analysis to assess System N functional redundancy. • Amino acid pH-dependent transport: SNAT5 transport is pH-sensitive — characterization under varied extracellular pH informs mechanism studies. EDITGENE recommends this model for researchers investigating glutamine transport biology, cancer cell metabolism, and System N transporter functions.

Yes. SNAT5 rescue experiments require attention to pH-dependence and sodium coupling: • Construct design: use a codon-modified SLC38A5 sequence with a small C-terminal tag (FLAG, HA). The 11 transmembrane SLC38 architecture must be preserved. • Transport-deficient rescue: substrate-binding pocket or sodium-binding residue mutations enable structure-function studies. • pH-dependent activity studies: SNAT5 transport is pH-sensitive; rescue lines should be characterized under defined pH conditions for consistent functional readouts. • Functional readout: rescue should restore sodium-dependent glutamine and asparagine uptake measured by radiolabel or LC-MS. HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation.