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A Library-based Approach Allows Systematic and Rapid Evaluation of Seed Region Length and Reveals Design Rules for Synthetic Bacterial Small RNAs

Overview
Journal iScience
Publisher Cell Press
Date 2024 Sep 16
PMID 39280619
Authors
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Abstract

All organisms must respond to environmental changes. In bacteria, small RNAs (sRNAs) are an important aspect of the regulation network underlying the adaptation to such changes. sRNAs base-pair with their target mRNAs, allowing rapid modulation of the proteome. This post-transcriptional regulation is usually facilitated by RNA chaperones, such as Hfq. sRNAs have a potential as synthetic regulators that can be modulated by rational design. In this study, we use a library-based approach and oxacillin susceptibility assays to investigate the importance of the seed region length for synthetic sRNAs based on RybB and SgrS scaffolds in . In the presence of Hfq we show that 12 nucleotides are sufficient for regulation. Furthermore, we observe a scaffold-specific Hfq-dependency and processing by RNase E. Our results provide information for design considerations of synthetic sRNAs in basic and applied research.

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References
1.
Lorenz R, Bernhart S, Honer Zu Siederdissen C, Tafer H, Flamm C, Stadler P . ViennaRNA Package 2.0. Algorithms Mol Biol. 2011; 6:26. PMC: 3319429. DOI: 10.1186/1748-7188-6-26. View

2.
Robertson H, Dunn J . Ribonucleic acid processing activity of Escherichia coli ribonuclease III. J Biol Chem. 1975; 250(8):3050-6. View

3.
Sprouffske K, Wagner A . Growthcurver: an R package for obtaining interpretable metrics from microbial growth curves. BMC Bioinformatics. 2016; 17:172. PMC: 4837600. DOI: 10.1186/s12859-016-1016-7. View

4.
Cho J, Yang D, Prabowo C, Ghiffary M, Han T, Choi K . Targeted and high-throughput gene knockdown in diverse bacteria using synthetic sRNAs. Nat Commun. 2023; 14(1):2359. PMC: 10126203. DOI: 10.1038/s41467-023-38119-y. View

5.
Popella L, Jung J, Do P, Hayward R, Barquist L, Vogel J . Comprehensive analysis of PNA-based antisense antibiotics targeting various essential genes in uropathogenic Escherichia coli. Nucleic Acids Res. 2022; 50(11):6435-6452. PMC: 9226493. DOI: 10.1093/nar/gkac362. View