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January 21st, 2025
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Jilin University
biochemistry
biorxiv

Transcriptome-wide identification of 5-methylcytosine by deaminase and reader protein-assisted sequencing

Zhou, J.Open in Google Scholar•Zhao, D.Open in Google Scholar•Li, J.Open in Google Scholar•Kong, D.Open in Google Scholar•Li, X.Open in Google Scholar•Zhang, R.Open in Google Scholar•Liang, Y.Open in Google Scholar•Gao, X.Open in Google Scholar•Qian, Y.Open in Google Scholar•Wang, D.Open in Google Scholaret al.

5-Methylcytosine (m5C) is one of the major post-transcriptional modifications in mRNA and is highly involved in the pathogenesis of various diseases. However, the capacity of existing assays for accurately and comprehensively transcriptome-wide m5C mapping still needs improvement. Here, we develop a detection method named DRAM (deaminase and reader protein assisted RNA methylation analysis), in which deaminases (APOBEC1 and TadA-8e) are fused with m5C reader proteins (ALYREF and YBX1) to identify the m5C sites through deamination events neighboring the methylation sites. This antibody-free and bisulfite-free approach provides transcriptome-wide editing regions which are highly overlapped with the publicly available BS-seq datasets and allows for a more stable and comprehensive identification of the m5C loci. In addition, DRAM system even supports ultra-low input RNA (10ng). We anticipate that the DRAM system could pave the way for uncovering further biological functions of m5C modifications.

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