SLC1A5 Knockout Cell Line (HCT 116)

The choice depends on whether you are studying SLC1A5 (ASCT2)'s role as the principal glutamine transporter or its emerging functions in cancer metabolism and mTORC1 signaling. The Knockout line is the standard tool for asking whether ASCT2 is required for glutamine uptake — ASCT2 is upregulated in many cancers and is the major route for glutamine entry in glutamine-addicted tumor cells, including colorectal cancer. Overexpression is useful for studying ASCT2 in cancer metabolism contexts or for assessing inhibitor specificity. For cancer metabolism research, the EDITGENE ASCT2 Knockout in HCT 116 is highly relevant — colorectal cancer cells frequently exhibit glutamine dependence, and ASCT2 is a validated cancer metabolism target. Rescue with wild-type or transport-deficient ASCT2 is the standard specificity control. The knockout serves as a critical genetic specificity tool for ASCT2 inhibitors (V-9302, IMD-0354 derivatives) and ASCT2-targeting therapeutic antibodies in development.

Primary applications: • Glutamine uptake: ³H-glutamine uptake assays to quantify ASCT2-dependent transport — ASCT2 is the principal glutamine uptake transporter in many cancers. • Cancer metabolism: glutaminolysis, anaplerotic flux into the TCA cycle, and proliferation under glutamine-restricted conditions. • mTORC1 signaling: phospho-S6K1 and phospho-4E-BP1 analysis given glutamine's role in mTORC1 activation. • ASCT2 inhibitor specificity: critical genetic control for V-9302 and other ASCT2-targeting compounds in cancer drug development. EDITGENE recommends this model for researchers investigating glutamine metabolism, cancer glutamine addiction, and ASCT2-targeted therapeutic development.

Yes. ASCT2 rescue experiments are well-established for cancer metabolism research: • Construct design: use a codon-modified SLC1A5 sequence with a small C-terminal tag (FLAG, HA). ASCT2 has 8 transmembrane domains in the canonical SLC1 architecture — preserve plasma membrane targeting determinants. • Transport-deficient rescue: substrate-binding pocket mutations enable distinguishing transport activity from non-transport functions. • V-9302-resistant rescue: ASCT2 inhibitor-resistant variants enable on-target validation of ASCT2-targeting compounds. • Functional readout: rescue should restore glutamine uptake activity and downstream mTORC1 signaling and proliferation phenotypes. HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation.