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RNA modifications (now frequently called epitranscriptomic RNA modifications) differ from unmodified parental counterpart (so A, C, G and U residues in primary RNA transcript) by their chemical structure and, in consequence, by their physicochemical properties (including base pairing properties), as well as by their chemical reactivity. These features allow mapping of RNA modifications by deep sequencing (both by second, NGS, and third, NNGS generations of sequencing technologies). Detection of modified residues is based either on specific RT signature during conversion of RNA sequence into cDNA, or on specific cleavage by chemical reagents allowing to create RT arrest. We developed and actively used different deep-sequencing approaches for mapping RNA modifications, namely 2′-O-methylations, some base methylation, pseudouridine, and others. Applications of these methods for epitranscriptomic analysis will be discussed.

Publié le juin 19, 2023