The AFMID Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, featuring targeted disruption of the AFMID (arylformamidase) gene. This model abrogates AFMID-catalyzed hydrolysis of N-formyl-L-kynurenine to kynurenine, a pivotal step in the kynurenine pathway of tryptophan metabolism. The heterogeneous polyclonal pool provides a robust platform for loss-of-function studies in an epithelial adenocarcinoma background.
HeLa cells are an immortalized human cervical adenocarcinoma line positive for HPV-18, extensively used in cancer biology and metabolic research. Their well-characterized signaling networks and metabolic profile make them amenable to investigating enzyme function and metabolic reprogramming. Incorporating AFMID disruption into this established cell model yields a stable tool for dissecting tryptophan catabolism within a carcinogenic context.
AFMID encodes arylformamidase, which functions downstream of indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO) to hydrolyze N-formylkynurenine into L-kynurenine. Kynurenine acts as a branchpoint for synthesis of quinolinic acid, NAD+, and immunomodulatory metabolites. CRISPR/Cas9-mediated AFMID disruption blocks this conversion, causing accumulation of N-formylkynurenine and reduced kynurenine, quinolinic acid, and NAD+ levels, thereby perturbing the IDO/TDO-AFMID-kynurenine axis and potentially impacting kynureninase and kynurenine 3-monooxygenase activities.
Within HeLa cells, the kynurenine pathway contributes to cancer cell proliferation, apoptosis resistance, and immune escape. AFMID knockout is anticipated to lower kynurenine availability, which may attenuate AhR-driven immunosuppressive programs and alter metabolic homeostasis. The HPV-18 positive adenocarcinoma origin provides a context to explore viral oncoprotein interplay with tryptophan metabolism. Being a polyclonal pool, this model captures population-level phenotypic effects, avoiding clonal selection bias and facilitating robust mechanistic studies.
Applications include quantitative LC-MS metabolite profiling, Western blot and RT-qPCR expression analysis of pathway enzymes, and functional assays such as cell proliferation, apoptosis, and migration. The model is suited for studying metabolic reprogramming, immune modulation, and drug sensitivity in cancer. This polyclonal knockout product is provided ready for immediate use. For further technical information, please contact Ascent Research.
