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Newly synthesized RNA Sequencing Characterizes Transcription Dynamics in Three Pluripotent States

By Rui Shao, Banushree Kumar, Katja Liedschreiber, Michael Maximilian Lidschreiber, Patrick Cramer, Simon J Elsässer

Posted 13 Jun 2021
bioRxiv DOI: 10.1101/2021.06.11.448016

Unique transcriptomes define naive, primed and paused pluripotent states in mouse embryonic stem cells. Here we perform transient transcriptome sequencing (TT-seq) to de novo define and quantify coding and non-coding transcription units (TUs) in different pluripotent states. We observe a global reduction of RNA synthesis, total RNA amount and turnover rates in ground state naive cells (2i) and paused pluripotency (mTORi). We demonstrate that elongation velocity can be reliably estimated from TT-seq nascent RNA and RNA polymerase II occupancy and observe a transcriptome-wide attenuation of elongation velocity in the two inhibitor-induced states. We also discover a relationship between elongation velocity and termination read-through distance. Our analysis suggests that steady-state transcriptomes in mouse ES cells are controlled predominantly on the level of RNA synthesis, and that signaling pathways governing different pluripotent states immediately control key parameters of transcription.

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