Ythdf2 Knockout Neuro-2a Cell Line

The Ythdf2 Knockout Neuro-2a Cell Line is a CRISPR/Cas9-edited mouse neuroblastoma cell line featuring targeted disruption of the Ythdf2 gene. This loss-of-function model enables stable ablation of YTHDF2 protein, providing a defined genetic background for investigating N6-methyladenosine (m6A)-dependent RNA regulation in a neuronal context. The cell line is suitable for dissecting m6A-mediated mRNA decay mechanisms and their impact on cell fate decisions.

The Neuro-2a host line, derived from an A/J strain mouse neuroblastoma, is widely used for studying neuronal differentiation and neurobiology. These cells can be differentiated with retinoic acid, leading to neurite outgrowth and expression of neuron-specific markers. Their tumorigenic nature also enables exploration of neuroblastoma pathogenesis, offering a model that bridges undifferentiated and differentiated states.

YTHDF2 is a cytoplasmic m6A reader that binds m6A-modified mRNAs and recruits the CCR4-NOT deadenylase complex to trigger transcript degradation. Its activity is regulated by upstream signals like hypoxia and HIF1A, and it controls the stability of downstream targets including MYC, NOTCH1, SOX2, and many neuronal mRNAs. YTHDF2 functions within an epitranscriptomic network involving the METTL3/METTL14 writer complex and the paralogs YTHDF1 and YTHDF3.

In Neuro-2a cells, Ythdf2 knockout disrupts the decay of m6A-marked transcripts, thereby stabilizing mRNAs that are normally rapidly turned over. This perturbation alters gene expression programs governing proliferation and neuronal differentiation, making the model ideal for studying how m6A dynamics influence neuroblastoma biology. The line is relevant to cancer research, given YTHDF2??s implication in glioblastoma and acute myeloid leukemia.

Researchers can employ this knockout line to investigate m6A modification in neuronal cells, analyze RNA decay in neuroblastoma, and dissect YTHDF2 function in differentiation and cancer. Compatible assays include Western blotting, RT-qPCR, RNA-seq, MeRIP-seq, proliferation assays, retinoic acid differentiation assays, and immunofluorescence. It also supports drug screening for m6A pathway inhibitors. For further details, contact Ascent Research.