GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE313nnn/GSE313744/primaryOK200GenomicsMus musculusNon-coding RNA profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE313744GEOGSEfalseThe MIME-seq technique allows to monitor the interaction of small non-coding RNAs with Argonaute proteins and their transfer to other cellsThe MIME-seq2.0 technique, developed by Mandlbauer et al., utilizes a chimeric HEN1 methyltransferase engineered to bind to Argonaute proteins (MIME enzyme) to protect the terminal sugar of small non-coding RNAs (sncRNAs) from oxidation, a reaction that inhibits RNA ligation and cloning. In this study, we confirm that the MIME-seq technique efficiently distinguish oxidized and non-oxidized sncRNAs using RNA sequencing and quantitative PCR. Upon transfection of the insulin-secreting cell line MIN6B1 with or without the MIME enzyme, we observed a significant loss in the detection of miRNAs following RNA oxidation, with a strong recovery rate in MIME-expressing cells. Comparison of the sncRNA profile in wild type and MIME-expressing cells demonstrated protection from oxidation not only of miRNAs, but also of some Y-RNA and tRNA-derived fragments. Immunoprecipitation with Ago2 antibodies confirmed that the protected Y-RNA and tRNA fragments bind to Argonaute proteins. We also used this system to track miRNA transfer between cells via extracellular vesicles (EVs). miRNAs released in EVs of MIME-expressing Jurkat T cells or C2C12 myotubes and delivered to MIN6B1 cells were protected from oxidation, enabling sensitive and specific detection of the transferred miRNAs in the receiving cells. Overall, the MIME-seq technique provides a powerful tool for analyzing RNA methylation and Argonaute binding in living cells and offers new possibilities for tracking RNA transfer across different cell types.2026/05/12GSE313744GSM9373406GSM9373409GSM9373407GSM9373408GSM9373412GSM9373413GSM9373410GSM937341134475313744Mus musculus[41844788]