Recently, a new rapid protein depletion system has been developed. In the past, RNAi knockdown systems were sometimes affected secondarily by long processing times. Combining this new high-speed replication system with comprehensive transcriptome analysis, we can uncover the events that initiate transcription in mammalian cells with unprecedented precision and depth. In doing so, we will identify and classify pseudo-transcription events that may regulate the expression of protein-coding genes in ways that are not yet understood. Although the molecular concepts involved in the initiation and termination of pseudo-transcription are already known, little is known about how these events affect the transcription of neighbouring genes. My project clarifies these examples and provides new information about the mechanism. Because of the fundamental questions being asked in this project and the mES cell lines involved, it is likely to have a broad impact on the fields of molecular and developmental biology. Therefore, the results of my research are valuable to the broader scientific community in molecular biology.
Indeed, so far I have first created a pipeline to determine new RNA transcription regions. This will help to determine not only the rapidly RNA-degrading transcripts that are the focus of this study, but also previously undiscovered transcripts induced by various stimuli (e.g. immune stimuli, osmotic pressure, etc.). The transcripts transcribed from uTSS were found to affect neighbouring mRNAs, and the biological significance of this is currently being further investigated. This finding suggests that single-lived RNAs, which are usually degraded, also have a function, and further developments in molecular biology are expected.
