NOV 28, 2017 07:00 AM PST

WEBINAR: Thiol-linked alkylation for the metabolic sequencing of RNA

SPONSORED BY: Lexogen
Speakers
  • Group Leader, Institute of Molecular Biotechnology (IMBA), Vienna, Austria
    Biography
      Dr. Stefan Ameres acquired his Master's degree in Biology at the Friedrich-Alexander-University Erlangen-Nuremberg in 2003, where he investigated during his Thesis the impact of DNA topology on transcriptional gene regulation. He then did his PhD Thesis in the lab of Renée Schröder at the Max F. Perutz Laboratories of the University of Vienna, where he performed the first in-depth enzymatic characterization of the human RNA interference effector complex. After a postdoc with Phillip Zamore at the University of Massachusetts Medical School in the US, where he focused on the small RNA-mediated gene regulation in flies and mammals he started his own lab in 2012 at the Institute of Molecular Biotechnology, IMBA, in Vienna, Austria. His lab investigates the mechanism and biology of post-transcriptional gene regulation, supported by the European research council through an ERC Starting Grant. Dr. Ameres is an elected Young Academy member of the Austrian Academy of Sciences and recently selected EMBO young investigator.

    Abstract:

    DATE: November 28, 2017
    TIME: 7:00AM PT, 10:00AM ET, 4:00PM CET

    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. We 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. We validated the method in mouse embryonic stem cells by showing that the RNA-polymerase-II-dependent transcriptional output scaled with Oct4/Sox2/Nanog-defined enhancer activity, and we provide quantitative and mechanistic evidence for transcript-specific RNA turnover mediated by post-transcriptional gene regulatory pathways initiated by microRNAs and N6-methyladenosine. SLAMseq facilitates the dissection of fundamental mechanisms that control gene expression in an accessible, cost-effective and scalable manner.


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