The YTHDF1 Knockout KYSE-140 Cell Line is a CRISPR/Cas9-edited knockout cell line engineered for targeted disruption of the YTHDF1 gene in human esophageal squamous epithelial cells. This model provides a powerful loss-of-function system for investigating the role of N6-methyladenosine (m6A) reader proteins in cancer biology. The cell line is offered as a ready-to-use in vitro tool, enabling researchers to study the functional consequences of abrogating YTHDF1-mediated translational regulation without the need for transient knockdown approaches.
The parental KYSE-140 cell line originates from a poorly differentiated esophageal squamous cell carcinoma (ESCC) derived from a Japanese male patient. As a well-characterized model of ESCC, KYSE-140 cells retain key histopathological and molecular features of aggressive squamous cell carcinoma, including maintained epithelial morphology and relevant oncogenic signaling networks. This cell line is widely employed in preclinical research to explore disease mechanisms and evaluate therapeutic strategies for esophageal cancer.
YTHDF1 functions as a cytoplasmic m6A reader protein that selectively recognizes m6A-modified mRNAs and recruits the translation initiation factor eIF3 to enhance cap-dependent translation. Its activity is regulated by m6A methyltransferase complexes containing METTL3, METTL14, and WTAP, and by demethylases such as FTO and ALKBH5. Upon binding m6A, YTHDF1 interacts with eIF3, poly(A)-binding protein PABPC1, and ribosomal subunits to promote efficient translation of target transcripts. Key downstream targets include CTNNB1 (??-catenin), MYC (c-Myc), SNAI1 (Snail), TCF7L2, and CCND1, which are critical effectors in pathways controlling proliferation, epithelial-mesenchymal transition (EMT), and metastasis. Through this mechanism, YTHDF1 integrates epitranscriptomic signals with oncogenic outputs, including Wnt/??-catenin, PI3K/AKT, and Hippo signaling networks.
In esophageal squamous cell carcinoma, YTHDF1 overexpression is frequently associated with enhanced translation of oncogenic mRNAs, driving tumor progression and poor prognosis. Disruption of YTHDF1 in the KYSE-140 background enables direct assessment of how m6A-dependent translational control contributes to ESCC pathogenesis. This knockout cell line serves as an ideal platform to investigate the dependency of ESCC cells on YTHDF1-mediated gene expression, and to evaluate the therapeutic potential of targeting the m6A machinery in this malignancy.
The YTHDF1 Knockout KYSE-140 Cell Line supports a broad range of functional studies. Typical applications include elucidation of m6A epitranscriptomic regulation in ESCC, systematic identification of YTHDF1 target mRNAs via RIP-seq or polysome profiling, and screening of small-molecule inhibitors targeting m6A writers, readers, or erasers. Compatible downstream assays include Western blotting and RT-qPCR for analyzing downstream protein and mRNA levels, m6A dot blot for global methylation assessment, cell proliferation assays (CCK-8/MTS), transwell migration and invasion assays, apoptosis assays (Annexin V/7-AAD), and in vivo xenograft tumor growth models. For additional information, please contact Ascent Research.
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