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Sequencing of polyA+ RNA from shCtrl and shMETTL8 HeLa for m3C site identification

GSE269470 Homo sapiens Expression profiling by high throughput sequencing; Other 6 samples Submitted 2026/07/17 Platform GPL24676
Summary
N3-methylcytidine (m3C), an RNA modification involving methylation of cytidine at the Watson-Crick face, has been identified in human tRNA, mitochondrial tRNA (mt-tRNA), and mRNA. In tRNA, it has been shown to play important roles in tRNA structure and translation. m3C remains understudied in mRNA due to its markedly lower levels, the lack of high-resolution techniques to reproducibly map its location, and under-characterization of its effector proteins on this RNA type. Additionally, METTL8 has been identified as a writer of m3C on mt-tRNA, but has also been shown to have isoforms with nuclear localization. Here, we apply a new approach which utilizes the high readthrough and mutation rates of Protoscript II (PSII) at m3C sites in combination with AlkB demethylation to map m3C at single-base resolution in polyA-purified RNA from HeLa cells either with METTL8-targeting shRNA or control shRNA to assess how the method works on mRNA and to investigate which, if any, of the sites change with METTL8 knockdown.
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Direct links to NCBI, no account and no request form: the whole study as GSE269470_RAW.tar, processed values as the series matrix, the supplementary file directory, and per-sample supplementary files for any of the 6 samples. Raw sequencing reads are also available from ENA.

Also filed as BioProject PRJNA1122093 and SRA study SRP512866. Searching any of these in the dataset finder brings you back here.

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