RHBG Knockout HCT 116 Cell Line

The choice depends on whether you are studying RHBG (Rh family B glycoprotein)'s role as an ammonium transporter or its specific functions in kidney epithelial ammonium handling and acid-base balance. The Knockout line is appropriate for asking whether RHBG is required for cellular ammonium transport — RHBG is principally expressed at basolateral membranes of kidney collecting duct cells, hepatic perivenous hepatocytes, and intestinal epithelium. Overexpression is useful for testing transport activity in heterologous systems or for studying disease-associated mutations. For ammonium transport research, the EDITGENE RHBG Knockout in HCT 116 enables study of intestinal ammonium handling — though renal ammonium transport research benefits from kidney epithelial models. RHCG paralog expression should be assessed given functional overlap. Rescue with wild-type or transport-deficient RHBG is the standard specificity control. The model is valuable for studying ammonium-related acid-base disorders and hepatic ammonia handling pathways.

Primary applications: • Ammonium transport: ¹⁴C-methylammonium (NH₄⁺ analog) uptake assays to quantify RHBG-dependent transport. • Acid-base handling: intracellular pH measurement under ammonium challenge to assess RHBG's contribution to pH regulation. • Hepatic ammonia handling: assessment of ammonia detoxification pathways in colorectal epithelial context. • Rh family paralog studies: RHCG and RHAG expression analysis to interpret RHBG-specific contributions. EDITGENE recommends this model for researchers investigating ammonium transporter biology, acid-base balance, and Rh family transport mechanisms.

Yes. RHBG rescue experiments require attention to membrane topology: • Construct design: use a codon-modified RHBG sequence with a small intracellular C-terminal tag (FLAG, HA). RHBG has 12 transmembrane domains — N-terminal tags must not disrupt topology. • Surface localization validation: confirm plasma membrane localization by cell surface biotinylation; basolateral targeting in epithelial cells requires specific signals. • Transport-deficient rescue: substrate-binding pocket mutations enable structure-function studies. • Functional readout: rescue should restore methylammonium uptake activity and ammonium-induced pH dynamics. HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation.