The ACER1 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population produced from the 786-O human renal cell adenocarcinoma cell line. Utilizing CRISPR/Cas9-mediated gene disruption, the alkaline ceramidase 1 (ACER1) locus is targeted to generate a heterogeneous pool of loss-of-function alleles, avoiding clonal isolation artifacts. This polyclonal knockout format provides a robust model system for investigating ACER1-dependent processes without the bias of single-cell-derived clones, making it suitable for population-level studies of sphingolipid biology and tumor cell behavior.
The parental 786-O cell line is a well-characterized clear cell renal cell carcinoma (ccRCC) model derived from a primary tumor of a 58-year-old male patient. 786-O cells harbor a biallelic loss-of-function mutation in the VHL tumor suppressor gene, resulting in constitutive stabilization of hypoxia-inducible factor (HIF) ?? subunits under normoxia. This VHL-mutant background drives pseudohypoxic gene expression programs, including upregulation of angiogenic factors and metabolic reprogramming, recapitulating hallmark features of ccRCC. The epithelial morphology and adherent growth characteristics of 786-O facilitate standardized assays for proliferation, migration, and drug sensitivity in a genetically defined renal cancer context.
ACER1 encodes an alkaline ceramidase that catalyzes the hydrolysis of ceramides to sphingosine, a key step in sphingolipid metabolism. Sphingosine is phosphorylated by sphingosine kinase 1 (SPHK1) to sphingosine-1-phosphate (S1P), which activates receptors S1PR1 and S1PR3. Downstream, S1P triggers AKT and ERK signaling, promoting cell survival and proliferation, while inhibiting protein kinase C (PKC). ACER1 is regulated by PPARG and NF-??B, and its activity intersects with ceramide synthase 1 and acid sphingomyelinase in the ceramide salvage pathway.
In the context of 786-O cells with constitutive HIF activation, ACER1 disruption is expected to alter the equilibrium between ceramide-mediated apoptosis and S1P-driven survival signals. The VHL mutation already sensitizes these cells to metabolic stress; ACER1 knockout may enhance ceramide accumulation and attenuate S1P production, potentially tipping the balance toward cell death or affecting migration and drug resistance. This engineered model thus enables precise dissection of how alkaline ceramidase activity modulates ccRCC cell fate and therapeutic vulnerability, without confounding clonal effects.
Typical applications include LC-MS ceramide profiling, sphingosine-1-phosphate ELISA, and confirmation of ACER1 knockout by Western blot or qPCR. Functional assays encompass apoptosis (Annexin V), proliferation (MTT/BrdU), and migration studies, along with phospho-flow cytometry for AKT, ERK, and PKC. These cells support research into renal cell carcinoma drug resistance and sphingolipid-related diseases such as melanoma, psoriasis, type 2 diabetes, and Parkinson’s disease. For further information, contact Ascent Research.
