Importantly, all the materials needed to perform this assay are commercially available, so the method is broadly accessible.

This has implications for the ability of pseudoknots to stabilize transcription pauses, which the Walter lab previously showed is possible (www.sciencedirect.com/science/arti...).

In addition to validating the method by assessing five RNA folding transitions in three RNA targets, we unexpectedly observed at least three pseudoknot base pairs forming within the RNA exit channel of RNAP.

This strategy is distinct from previous methods (Cotranscriptional SHAPE-Seq, SPET-Seq, TECprobe-VL), which measure cotranscriptionally folded RNA, but not cotranscriptional RNA folding - in our new approach, we directly assess how RNA structure changes as RNAP transcribes downstream.

In this study (www.nature.com/articles/s41...) we used a reversible transcription roadblocking strategy developed by the Lafontaine lab to sequentially position E. coli RNAP at two or three positions within a DNA template so that we can take fractions of the reaction for chemical probing.

Actually, I was off on the year of Opiate, so only Fear Inoculum counts (better album IMHO)

Have you watched Twin Peaks?

More or less sense than using pipes as the primary means of transit?

Apparently since the beginning!

Thanks!

We designed the TECdisplay platform to be broadly accessible and easy to use, so our hope is that this approach democratizes the development and application of high-throughput RNA biochemical assays.

We applied this to a ZTP riboswitch variant library to investigate the mechanisms by which variants that match the consensus sequence and structure misfold. Panel c illustrates the ZMP-mediated antitermination response for the 32,768 variants in the library!

Our first TECdisplay assay measures transcription termination efficiency: TECs are tethered to a bead by the nascent RNA and DNA is released from the bead if transcription is terminated, but retained on the bead if RNA polymerase bypasses the terminator.