SLC36A1 Knockout HCT116 Cell Line

The choice depends on whether you are studying SLC36A1 (PAT1)'s role as a proton-coupled amino acid transporter or its specific contributions to intestinal absorption of small amino acids and amino acid-like drugs. The Knockout line is the standard tool for asking whether PAT1 is required for proton-dependent uptake of glycine, proline, alanine, and γ-aminobutyric acid (GABA) — particularly relevant in HCT 116, which has intestinal epithelial origins. Overexpression is useful for testing transport activity in heterologous systems and for assessing drug-substrate interactions. For PAT1 research, the EDITGENE Knockout in HCT 116 is highly informative for intestinal amino acid absorption and oral drug bioavailability studies — PAT1 contributes to apical uptake of certain GABA-related drugs (vigabatrin, gabapentin analogs) and δ-aminolevulinic acid (5-ALA used in photodynamic therapy). Rescue with wild-type or transport-deficient PAT1 enables structure-function studies, and is a critical specificity control for PAT1-mediated drug uptake research.

Primary applications: • Proton-coupled amino acid uptake: ³H-glycine, ³H-proline, or ³H-GABA uptake assays under varied extracellular pH conditions to characterize PAT1 transport activity. • Drug uptake studies: 5-ALA (photodynamic therapy), vigabatrin, and gabapentin analog uptake assays — these drugs are substrates of PAT1 for intestinal absorption. • Intestinal absorption modeling: PAT1-dependent oral bioavailability assessment for drug development in a colorectal epithelial context. • Paralog studies: SLC36A2/SLC36A3/SLC36A4 expression analysis to characterize PAT family functional overlap. EDITGENE recommends this model for researchers investigating proton-coupled amino acid transport, oral drug bioavailability, and amino acid-mimetic drug development.

Yes. PAT1 rescue experiments require attention to proton-coupling and pH-dependence: • Construct design: use a codon-modified SLC36A1 sequence with a small C-terminal tag (FLAG, HA). PAT1 has 11 transmembrane domains — N-terminal tags must not disrupt topology. • Transport-deficient rescue: conserved residue mutations in proton-binding sites or substrate-binding pockets enable structure-function studies. • pH-dependence: PAT1 transport is highly pH-sensitive — rescue assays should be performed under controlled extracellular pH conditions (typically pH 5.5-6.5 for optimal activity). • Functional readout: rescue should restore proton-coupled amino acid uptake measured by ³H-glycine or related substrate uptake. HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation for intestinal-relevant transport studies.