The ABHD12 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HCT 116 human colorectal carcinoma cell line. Targeted disruption of the ABHD12 gene eliminates expression of the serine hydrolase that hydrolyzes lysophosphatidylserine (lyso-PS), providing a loss-of-function model for studying lyso-PS signaling. The polyclonal format retains genetic diversity, suitable for pooled screens and functional assays.
HCT 116 is an epithelial colorectal carcinoma cell line harboring an MLH1 mutation, causing microsatellite instability, and a KRAS G13D activating mutation. These features make it a standard model for colorectal cancer research, including studies of tumor progression, drug sensitivity, and DNA repair. The hypermutable background and oncogenic KRAS signaling provide a relevant platform for investigating lipid-mediated pathways in cancer.
ABHD12 functions as a critical regulator of immune signaling by hydrolyzing lyso-PS, thereby terminating activation of the GPR34 receptor. Its expression is upregulated by TLR ligands such as LPS and cytokines like IL-1?? and TNF-?? via NF-??B and AP-1 transcription factors. Downstream, ABHD12 dampens GPR34/G??i/o-mediated signals, reducing MAPK/ERK and PI3K/AKT pathway activity and suppressing secretion of IL-10 and TGF-??. This positions ABHD12 at the intersection of innate immunity, lipid metabolism, and inflammatory control.
In HCT 116 cells, ABHD12 knockout enables dissection of lyso-PS metabolism in colorectal cancer. The mutant KRAS and MSI background may influence how accumulated lyso-PS alters GPR34 signaling, impacting cell proliferation, migration, and immune interactions. This model is valuable for studying tumor-microenvironment crosstalk, particularly macrophage phagocytosis and cytokine modulation, and for exploring ABHD12 as a potential target in cancer therapy.
Applications include high-throughput drug screening, CRISPR screen controls, GPR34 reporter assays, and phagocytosis assays. ABHD12 ablation can be confirmed by western blotting, while LC-MS quantifies lyso-PS and ELISA measures cytokines like IL-10 and TGF-??. Additional uses include proliferation and migration assays, RNA-seq transcriptomics, and disease modeling for PHARC syndrome. For more information, contact Ascent Research.
