The Fto Knockout H9c2(2-1) Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the H9c2(2-1) rat cardiomyoblast cell line, featuring targeted disruption of the Fto gene. This cell line serves as a loss-of-function model, abrogating expression of the FTO RNA N6-methyladenosine (m6A) demethylase to enable investigation of m6A-dependent gene regulation in a cardiac cell context.
The parental H9c2(2-1) line, a subclone of H9c2 cardiomyoblasts originally isolated from embryonic BDIX rat heart tissue, is a well-established in vitro model for cardiac muscle biology. These precursor cells can differentiate into cardiomyocyte-like cells and are widely employed for studying cardiac metabolism, hypertrophy, differentiation, and stress responses.
FTO mediates oxidative demethylation of m6A on mRNA, thereby regulating transcript stability, splicing, and translation. FTO is regulated by upstream factors such as the energy sensor AMPK, insulin, glucocorticoids, and transcription factors PPARG and CEBPA, and interacts with core m6A methyltransferase complex components METTL3, METTL14, and WTAP. Its demethylase activity modulates binding of YTHDF1, YTHDF2, and YTHDF3 reader proteins, influencing downstream targets including LEP, ADIPOQ, UCP1, and PPARG itself. Through these interactions, FTO integrates m6A modification with energy homeostasis, adipogenesis, insulin signaling, and AMPK/mTOR pathways.
In H9c2(2-1) cardiomyoblasts, FTO knockout enables direct dissection of m6A epitranscriptomic mechanisms in cardiac contexts. Loss of FTO is expected to alter m6A modification patterns on transcripts encoding proteins involved in metabolism, contractile function, and stress adaptation, providing a model to study how RNA methylation impacts cardiomyocyte differentiation and cardiac function. This system is particularly useful for exploring the interplay between FTO-regulated pathways??such as insulin and AMPK signaling??and heart disease, and for investigating the role of m6A in the cardiac response to obesity-associated signals.
The Fto Knockout H9c2(2-1) Cell Line supports a wide range of research applications, including epitranscriptomic profiling by m6A RNA immunoprecipitation and sequencing (MeRIP-seq), gene expression analysis via RT-qPCR and Western blotting, and phenotypic assessments using flow cytometry for differentiation markers, immunofluorescence localization, metabolic flux assays (e.g., Seahorse), apoptosis assays, and scratch wound migration assays. It is also amenable to chemical screening for FTO inhibitors and to functional validation of m6A-modified transcripts in cardiac metabolism and disease models. For additional information, please contact Ascent Research.
