GPR65 Knockout Cell Line (HEK293)

The choice depends on whether you are studying GPR65 (TDAG8)'s role as a proton-sensing GPCR or its functions in immune cell biology and inflammatory bowel disease. The Knockout line is the standard tool for asking whether GPR65 is required for these processes — GPR65 is one of four proton-sensing GPCRs (with GPR4, GPR68/OGR1, GPR132/G2A) that respond to extracellular acidification through histidine residue protonation, signaling through Gs (cAMP) and other G proteins. Overexpression is useful for studying GPR65 in heterologous expression contexts. For acid-sensing GPCR research, the EDITGENE GPR65 Knockout in HEK293 enables study of proton-sensing biology — GPR65 variants are associated with inflammatory bowel disease susceptibility (GWAS). Rescue with wild-type or histidine-mutant GPR65 enables structure-function studies of pH sensing. The knockout is valuable for studying tumor microenvironment acidosis sensing and emerging proton-sensing GPCR-targeted therapeutics.

Primary applications: • Proton-sensing signaling: cAMP measurement and downstream Gs/Gq signaling following extracellular acidification (pH 6.5-7.4) in GPR65-null cells. • IBD susceptibility studies: rescue with GPR65 disease-associated variants (e.g., I231L) for genotype-function studies of inflammatory bowel disease. • Tumor microenvironment acidosis: in heterologous tumor-relevant contexts, characterization of GPR65's role in acidosis-sensing. • Proton-sensing GPCR comparative studies: GPR4, GPR68 (OGR1), GPR132 expression analysis for family-wide pH-sensing studies. EDITGENE recommends this model for researchers investigating proton-sensing GPCR biology, IBD susceptibility, and tumor microenvironment acidosis signaling.

Yes. GPR65 rescue experiments require attention to proton-sensing histidine residues: • Construct design: use a codon-modified GPR65 sequence with a small intracellular C-terminal tag (FLAG, HA). GPR65 is a seven-transmembrane GPCR with key extracellular histidine residues serving as pH sensors — preserve all elements. • Histidine-mutant rescue: extracellular histidine residue mutations (H6Q, H8Q) abolish pH-sensing and serve as the standard specificity control. • IBD risk variant rescue: I231L and other IBD-associated GPR65 variants enable disease genotype-function studies. • Functional readout: rescue should restore acidification-induced cAMP elevation and downstream Gs signaling. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.