Description

This product is a CRISPR/Cas9-edited Mettl3 knockout cell line derived from Neuro-2a cells. Through CRISPR/Cas9-mediated gene disruption, the Mettl3 locus has been targeted to eliminate expression of the catalytic subunit of the N6-methyladenosine (m6A) methyltransferase complex, providing a clean loss-of-function model for epitranscriptomic research.

Neuro-2a is a mouse neural crest-derived neuroblastoma line, a subclone of C1300, extensively utilized as a neuronal model for studying differentiation and neurite outgrowth. Serum starvation induces these cells to extend neurites and express neuronal markers, making them ideal for investigating molecular mechanisms of neurogenesis.

Mettl3 associates with METTL14, WTAP, VIRMA, RBM15, and ZC3H13 to form the m6A writer complex, which catalyzes co-transcriptional deposition of m6A on mRNA. This modification influences RNA stability, splicing, and translation. Mettl3 activity is modulated by upstream regulators including WTAP, c-Myc, and KMT2A, and its catalytic output directly methylates transcripts such as Neurog2, Mapt, Bdnf, and Sox2 mRNAs. Recognition of m6A marks by reader proteins (YTHDF1, YTHDF2, YTHDF3) and removal by erasers (FTO, ALKBH5) complete a dynamic regulatory network.

In the Neuro-2a neuronal context, Mettl3-mediated m6A methylation is critical for the post-transcriptional control of genes driving neuronal differentiation. Knockout of Mettl3 destabilizes key mRNAs, including Neurog2 and Mapt, resulting in impaired neurite outgrowth and neuronal maturation. This cell line thus enables dissection of epitranscriptomic layers in neurodevelopment and serves as a model for m6A-related disorders such as neurodevelopmental syndromes and glioblastoma.

Researchers can apply this knockout line in m6A MeRIP-seq to map transcriptome-wide methylation changes, RNA-seq for differential expression analysis, and RT-qPCR for target gene validation. Functional assays include neurite outgrowth measurement, immunofluorescence staining for neuronal markers, and metabolic profiling. Additionally, the line is valuable for mechanistic studies of mRNA decay and translation, rescue experiments with wild-type or mutant Mettl3, and high-throughput screening of molecules targeting the m6A pathway. For further assistance, please contact Ascent Research.