The ACSL4 Knockout TE1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt ACSL4 gene expression in the TE1 human esophageal squamous carcinoma background. This pooled knockout model provides a versatile loss-of-function system for interrogating ACSL4-dependent cellular processes without clonal selection pressures. The polyclonal format preserves heterogeneous gene editing outcomes, enabling studies of population-level responses relevant to tumor heterogeneity.
The TE1 cell line is an established epithelial model derived from a human esophageal squamous cell carcinoma. TE1 cells exhibit characteristic features of ESCC, including dysregulated lipid metabolism and susceptibility to ferroptotic cell death. As a widely utilized platform in cancer research, TE1 cells allow investigation of oncogenic signaling, drug resistance, and lipid-mediated cell death mechanisms.
ACSL4 (acyl-CoA synthetase long-chain family member 4) catalyzes the activation of long-chain polyunsaturated fatty acids, preferentially utilizing arachidonic acid, to generate PUFA-CoA esters. These products are esterified into membrane phospholipids by LPCAT3, rendering membranes susceptible to oxidation by lipoxygenases (LOXs). Subsequent lipid peroxidation drives ferroptosis, an iron-dependent regulated necrosis executed by the collapse of the lipid peroxide-sensing system involving GPX4. ACSL4 expression is positively regulated by transcription factors SP1, SREBP1, and PPAR??, and is downstream of PI3K/AKT signaling. Through this axis, ACSL4 functions as a critical node linking fatty acid metabolism to ferroptosis execution.
In the context of esophageal squamous cell carcinoma, ACSL4 governs sensitivity to ferroptosis-inducing agents. The TE1 knockout model permits dissection of how loss of ACSL4-mediated PUFA activation alters lipidomic profiles, reduces lipid peroxidation, and confers resistance to ferroptosis. This system is valuable for exploring ferroptosis evasion as a therapeutic resistance mechanism and for screening compounds that bypass ACSL4 dependency to restore cell death in ESCC.
Researchers can employ this polyclonal knockout product for ferroptosis induction assays (e.g., erastin, RSL3 treatment), lipid peroxidation quantification using C11-BODIPY or malondialdehyde assays, western blotting of key pathway components such as GPX4 and LOXs, and RNA-seq?Cbased transcriptomic profiling. Metabolic assays assessing arachidonoyl-CoA production and phospholipid remodeling further characterize the functional consequences of ACSL4 loss. Additionally, migration and drug sensitivity assays help delineate ACSL4??s role in ESCC progression and treatment response. For further technical information or to inquire about custom projects, please contact Ascent Research.
