Broccoli Aptamer Allows Quantitative Transcription Regulation Studies in Vitro

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2024 Jun 13

PMID 38870160

Authors

Amanda van der Sijs

Thomas Visser

Pepijn Moerman

Gert Folkers

Willem Kegel

Affiliations

Soon will be listed here.

Abstract

Quantitative transcription regulation studies in vivo and in vitro often make use of reporter proteins. Here we show that using Broccoli aptamers, quantitative study of transcription in various regulatory scenarios is possible without a translational step. To explore the method we studied several regulatory scenarios that we analyzed using thermodynamic occupancy-based models, and found excellent agreement with previous studies. In the next step we show that non-coding DNA can have a dramatic effect on the level of transcription, similar to the influence of the lac repressor with a strong affinity to operator sites. Finally, we point out the limitations of the method in terms of delay times coupled to the folding of the aptamer. We conclude that the Broccoli aptamer is suitable for quantitative transcription measurements.

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