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This week Dr. Stefan Ameres, a group leader at the Institute of Molecular Biotechnology (IMBA) in Vienna, Austria gave a webinar talk about thiol-linked alkylation for the metabolic sequencing of RNA. This technology provides a basis for Lexogen’s family of SLAMseq kits, launched on September 25, 2017 after its official publication in the Nature Methods journal (Herzog, V.A. et al., DOI: 10.1038/nmeth.4435).
In this webinar Dr. Ameres explained that gene expression profiling by high-throughput sequencing reveals qualitative and quantitative changes in RNA species at steady state but obscures the intracellular dynamics of RNA transcription, processing and decay.
His laboratory developed thiol(SH)-linked alkylation for the metabolic sequencing of RNA (SLAMseq), an orthogonal-chemistry-based RNA sequencing technology that detects 4-thiouridine (s4U) incorporation in RNA species at single-nucleotide resolution. In combination with well-established metabolic RNA labeling protocols and coupled to standard, low-input, high-throughput RNA sequencing methods, SLAMseq enabled rapid access to RNA-polymerase-II-dependent gene expression dynamics in the context of total RNA.
The method was validated in mouse embryonic stem cells by showing that the RNA-polymerase-II-dependent transcriptional output scaled with Oct4/Sox2/Nanog-defined enhancer activity. Quantitative and mechanistic evidence for transcript-specific RNA turnover mediated by post-transcriptional gene regulatory pathways initiated by microRNAs and N6-methyladenosine was provided.
Dr. Ameres confirmed in his talk that SLAMseq facilitates the dissection of fundamental mechanisms that control gene expression in an accessible, cost-effective and scalable manner.
