Kurzfassung

Post-transcriptional diversification of the genome output massively contributes to accomplish the demands of a cell to execute biological programs to establish and maintain the integrity of complex multicellular organisms. Deregulation of this process can cause developmental defects, and aberrant transcriptome diversification is widespread in numerous disease processes including cancer. Here we establish a method based on deep-sequencing, which allows interrogating the dynamics of...Post-transcriptional diversification of the genome output massively contributes to accomplish the demands of a cell to execute biological programs to establish and maintain the integrity of complex multicellular organisms. Deregulation of this process can cause developmental defects, and aberrant transcriptome diversification is widespread in numerous disease processes including cancer. Here we establish a method based on deep-sequencing, which allows interrogating the dynamics of transcriptome 3’end diversity in a highly multiplexed fashion (TREND-seq).
 
As a proof of concept, we analyzed the transcriptome diversity in response to individual depletion of more than 170 potential executing regulators by RNAi in human neuroblastoma. We demonstrate that this protocol reliably works with minimal amounts of RNA (< 100 ng) following a straight-forward fast and scalable workflow. In our experimental model system we observe a massive deregulation of transcriptome 3’end diversity during tumorigenesis, which - interestingly - affects numerous oncogenes and tumorsuppressors.

TREND-seq thus represents a powerful tool for elucidating the dynamics of transcriptome 3’ end diversity in a highly multiplexed format in various (patho-)physiological processes.» weiterlesen» einklappen