FTO Knockout Cell Line (HCT 116)

The choice depends on whether you are studying FTO (fat mass and obesity-associated)'s role as the principal m6A (N6-methyladenosine) RNA demethylase or modeling its functions in human colorectal cancer biology and metabolic disease. The Knockout line is the standard tool for asking whether FTO is required for these processes — FTO is a 2OG/Fe(II)-dependent dioxygenase that removes m6A from mRNA (and m1A from tRNA, m6Am from cap structures), making it one of two principal m6A 'erasers' (with ALKBH5); FTO common variants are the strongest human genetic association with obesity (T allele of rs9939609 increases obesity risk). Overexpression is useful for studying FTO in heterologous expression contexts. For colorectal cancer and m6A epitranscriptomics research, the EDITGENE FTO Knockout in HCT 116 is highly relevant — HCT 116 is a colorectal cancer cell line, and FTO has emerged as an oncogene in multiple cancers including colorectal cancer where m6A epitranscriptomic regulation drives proliferation. This product complements the parallel FTO Knockout in MAC-T (bovine mammary, also available) for human cancer versus agricultural genomics studies. Rescue with wild-type or catalytically-dead (H231A) FTO is the standard specificity control. The knockout is a critical specificity tool for FTO inhibitors (FB23-2, meclofenamic acid, R-2HG) and m6A-targeted cancer therapeutics.

Primary applications: • m6A epitranscriptomics: m6A-RIP-seq, MeRIP-seq, or SCARLET analysis of mRNA m6A modifications in FTO-null versus rescued colorectal cancer cells. • Colorectal cancer biology: proliferation, migration, and apoptosis assays given FTO's emerging oncogenic role. • Cross-species comparison: parallel analysis with FTO Knockout in MAC-T (bovine, also available) for human cancer versus agricultural genomics studies. • FTO inhibitor specificity: critical genetic control for FB23-2, meclofenamic acid, R-2HG, and other FTO-targeting compounds in cancer and obesity drug development. EDITGENE recommends this HCT 116-based model for human cancer m6A research; the parallel FTO Knockout in MAC-T (also available) is preferred for bovine lactation and agricultural genomics studies.

Yes. FTO rescue experiments require attention to dioxygenase architecture: • Construct design: use a codon-modified FTO sequence with a small C-terminal tag (FLAG, HA). FTO has N-terminal AlkB-like dioxygenase domain and C-terminal β-helix domain — preserve all elements. • Catalytically-dead rescue: H231A or D233A active site Fe(II)-coordinating residue mutations abolish demethylase activity and serve as the standard specificity control. • Functional readout: rescue should restore m6A demethylase activity measured by global mRNA m6A levels (LC-MS) or m6A-RIP-seq. HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation.