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High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes

High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes

Published on: March 3, 2015

Screening assays to identify artificial glmS ribozyme activators.

Christina E Lünse¹, Günter Mayer

¹Life and Medical Sciences Institute, University of Bonn, Bonn, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|December 10, 2013

Summary

Riboswitches regulate essential bacterial genes and are potential drug targets. We developed high-throughput assays to screen for molecules that activate bacterial riboswitches, aiding drug discovery.

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Area of Science:

Molecular Biology
RNA Biology
Drug Discovery

Background:

Riboswitches are regulatory RNA elements controlling gene expression in bacteria.
Their role in essential gene regulation makes them attractive targets for antimicrobial drug development.
Developing high-throughput screening (HTS) assays is crucial for identifying novel riboswitch-targeting compounds.

Purpose of the Study:

To establish robust, high-throughput compatible assays for screening riboswitch activators.
To identify artificial molecules capable of activating bacterial riboswitches.

Main Methods:

Developed a fluorescence-based screening assay using a minimal Bacillus subtilis glmS riboswitch motif.
Employed a metabolite-induced self-cleavage assay with the full-length Staphylococcus aureus glmS ribozyme.
Utilized these assays for high-throughput screening of compound libraries.

Main Results:

Successfully established two distinct HTS-compatible assays for riboswitch activation screening.
Demonstrated the utility of these assays in identifying potential riboswitch-activating molecules.
Provided a foundation for discovering novel antibacterial agents targeting riboswitches.

Conclusions:

The developed assays are effective for high-throughput screening of riboswitch activators.
This work facilitates the discovery of new therapeutic strategies targeting bacterial gene regulation.
The identified molecules represent potential leads for novel antibiotic development.