The ACSL4 Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ACSL4 gene in the human bladder cancer cell line UM-UC-3. This loss-of-function model enables investigation of ACSL4-dependent processes without any presupposed mutation type, providing a versatile tool for functional genomics.
UM-UC-3 is a well-characterized epithelial cell line derived from a urinary bladder transitional cell carcinoma in a male patient. These adherent cells exhibit typical carcinoma characteristics and serve as a relevant model for studying bladder cancer biology, including tumor progression, metastasis, and therapeutic responses.
ACSL4 encodes an acyl-CoA synthetase that preferentially activates polyunsaturated fatty acids (PUFAs) by conjugating them with coenzyme A. This reaction facilitates the incorporation of PUFAs into membrane phospholipids, a process mediated by downstream targets such as phospholipid remodeling enzymes. ACSL4 activity is transcriptionally regulated by upstream factors SREBP1c and HIF-1??, and its product line interfaces with interacting partners GPX4 and LPCAT3. The resulting PUFA-enriched membranes are susceptible to lipid peroxidation, driven by iron-dependent LOX enzymes and lipid hydroperoxide accumulation, ultimately promoting ferroptotic cell death. Knockout of ACSL4 disrupts this cascade, diminishing ferroptosis sensitivity and altering cellular lipidome composition.
In the context of the UM-UC-3 bladder carcinoma background, ACSL4 polyclonal knockout cells offer a platform to dissect the interplay between lipid metabolism and ferroptosis susceptibility in cancer. Bladder cancer studies increasingly implicate ferroptosis evasion as a mechanism of drug resistance, and this model enables direct assessment of how loss of ACSL4 function impacts cell viability under ferroptosis-inducing agents such as erastin. Such investigations can reveal vulnerabilities or adaptive responses specific to urothelial carcinoma.
Research applications of this polyclonal knockout population include mechanistic studies of ferroptosis regulation, lipidomic profiling by LC-MS, and functional validation of ACSL4 in cancer cell growth and therapy resistance. Representative assays like Western blotting, RT-qPCR, lipid peroxidation assays, and cellular viability measurements following ferroptosis induction are highly compatible with this model. Immunofluorescence and CRISPR functional validation can further confirm gene disruption and phenotypic outcomes. Researchers are encouraged to contact Ascent Research for detailed technical specifications, lot-specific quality control data, and personalized experimental guidance.
