This CRISPR/Cas9-edited polyclonal knockout cell population is derived from the KYSE-30 human esophageal squamous cell carcinoma cell line with targeted disruption of the AIFM2 gene. The polyclonal nature provides a heterogeneous pool of gene-edited cells, enabling study of AIFM2 loss-of-function without clonal selection bias. This model serves as a tool for investigating AIFM2 roles in apoptosis and cancer biology.
The parental KYSE-30 cell line originates from a well-differentiated esophageal squamous cell carcinoma of a 64-year-old male. These epithelial cells retain tumor characteristics and are a standard model for esophageal cancer research. KYSE-30 cells are suitable for studying tumor biology and drug responses, and the AIFM2 knockout allows direct investigation of AIFM2-dependent mechanisms in this context.
AIFM2 (apoptosis-inducing factor 2) is a mitochondrial flavoprotein oxidoreductase that mediates caspase-independent apoptosis. Under stress such as DNA damage or hypoxia, AIFM2 is activated downstream of TP53 and E2F1, translocating to the nucleus to interact with DNA, HSPA1A, and PPIA. This promotes chromatin condensation and DNA fragmentation via endonuclease activation, bypassing caspase cascades. AIFM2??s role in mitochondrial dysfunction-mediated cell death is linked to mitochondrial outer membrane permeabilization. Knockout of AIFM2 disrupts this alternative death pathway, allowing dissection of its contributions to stress responses.
In KYSE-30 esophageal cancer cells, AIFM2 loss may attenuate caspase-independent apoptosis, potentially enhancing survival and chemoresistance. This is relevant for esophageal squamous cell carcinoma, where apoptosis evasion drives tumor progression. Comparing knockout and wild-type cells enables delineation of AIFM2??s role in tumorigenicity and drug sensitivity, and its impact on mitochondrial dysfunction-mediated death.
The AIFM2 knockout KYSE-30 polyclonal cells are applicable for western blotting, RT-qPCR, and RNA-seq to confirm disruption and transcriptomic changes. Apoptosis assays (annexin V, TUNEL), colony formation, and drug sensitivity testing evaluate functional consequences. Chromatin condensation assays and immunofluorescence for AIFM2 localization further characterize the phenotype. These cells support studies on caspase-independent apoptosis, chemoresistance, and signaling interactions with TP53 and HSPA1A. Contact Ascent Research for further details.
