ACSL4 Knockout 786-O Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ACSL4 gene in the human 786-O epithelial cell line. This polyclonal pool contains a heterogeneous mixture of gene disruptions introduced by CRISPR/Cas9-mediated genome editing, creating a loss-of-function model for ACSL4-dependent pathways without requiring single-cell cloning. The product provides researchers with a physiologically relevant system to investigate ACSL4 biology in a defined renal carcinoma background, facilitating studies of ferroptosis, lipid metabolism, and oncogenic signaling.
Derived from a primary clear cell renal cell carcinoma, the 786-O host line is widely used in cancer research due to its well-characterized VHL tumor suppressor mutation. This VHL deficiency leads to constitutive stabilization of hypoxia-inducible factors (HIFs), driving metabolic reprogramming, enhanced lipid droplet accumulation, and altered redox homeostasis. The epithelial origin and tumorigenic properties of 786-O cells make them an appropriate model for exploring the intersection of metabolic vulnerabilities and ferroptotic cell death mechanisms in kidney cancer.
ACSL4 encodes a long-chain fatty acyl-CoA synthetase that preferentially activates polyunsaturated fatty acids, such as arachidonic acid, into acyl-CoA esters. These activated fatty acids are subsequently incorporated into membrane phospholipids via LPCAT3, providing substrates for lipoxygenase-mediated peroxidation. ACSL4 expression is transcriptionally regulated by SREBP1 and PPAR??, and its activity is modulated by upstream signals including TFEB and hormonal cues. In the ferroptosis execution network, ACSL4 operates downstream of system Xc- (SLC7A11/SLC3A2) and upstream of ALOX5-dependent lipid peroxidation, with the generated phospholipid hydroperoxides normally neutralized by GPX4. Thus, ACSL4 serves as a central node linking fatty acid metabolism to oxidative cell death.
In the 786-O background, endogenous ACSL4 contributes to the generation of pro-ferroptotic lipid species, and its disruption is expected to alter sensitivity to ferroptosis-inducing agents such as erastin and RSL3. The VHL-mutant context of these cells further sensitizes them to metabolic perturbations, making this polyclonal knockout population a valuable tool for dissecting ACSL4??s role in lipid-mediated cell death and therapy resistance. Researchers can leverage this model to explore how loss of ACSL4 affects lipid peroxidation dynamics, phospholipid remodeling, and clonogenic survival under oxidative stress.
This product enables a wide range of experimental applications relevant to ferroptosis biology, metabolic reprogramming, and drug sensitivity profiling. Representative assays include Western blotting for ACSL4 depletion, C11-BODIPY staining for lipid peroxidation quantification, and cell viability assessments following treatment with canonical ferroptosis inducers. Additional downstream analyses such as RT-qPCR, immunofluorescence for lipid droplet morphology, and acyl-CoA synthetase activity measurements are highly compatible. The polyclonal knockout cells are particularly suited for pooled screening approaches and mechanistic studies where clonal homogeneity is not required. For detailed technical specifications and ordering information, please contact Ascent Research.
