Webb, Rebecca J., Roberts, Alexandra A., Rush, Catherine, Skerratt, Lee F., Tizard, Mark L., and Berger, Lee (2024) Small Interfering RNA Mediated Messenger RNA Knockdown in the Amphibian Pathogen Batrachochytrium dendrobatidis. Journal of Basic Microbiology, 64 (8).

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Abstract

RNA interference (RNAi) has not been tested in the pandemic amphibian pathogen, Batrachochytrium dendrobatidis, but developing this technology could be useful to elucidate virulence mechanisms, identify therapeutic targets, and may present a novel antifungal treatment option for chytridiomycosis. To manipulate and decipher gene function, rationally designed small interfering RNA (siRNA) can initiate the destruction of homologous messenger RNA (mRNA), resulting in the “knockdown” of target gene expression. Here, we investigate whether siRNA can be used to manipulate gene expression in B. dendrobatidis via RNAi using differing siRNA strategies to target genes involved in glutathione and ornithine synthesis. To determine the extent and duration of mRNA knockdown, target mRNA levels were monitored for 24–48 h after delivery of siRNA targeting glutamate–cysteine ligase, with a maximum of ~56% reduction in target transcripts occurring at 36 h. A second siRNA design targeting glutamate-cysteine ligase also resulted in ~53% knockdown at this time point. siRNA directed toward a different gene target, ornithine decarboxylase, achieved 17% reduction in target transcripts. Although no phenotypic effects were observed, these results suggest that RNAi is possible in B. dendrobatidis, and that gene expression can be manipulated in this pathogen. We outline ideas for further optimization steps to increase knockdown efficiency to better harness RNAi techniques for control of B. dendrobatidis.

Item ID:	85693
Item Type:	Article (Research - C1)
ISSN:	1521-4028
Copyright Information:	This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. © 2024 The Author(s). Journal of Basic Microbiology published by Wiley‐VCH GmbH.
Funders:	Australian Research Council (ARC)
Projects and Grants:	ARC DP220101361, ARC FT190100462
Date Deposited:	03 Jun 2025 00:53
FoR Codes:	31 BIOLOGICAL SCIENCES > 3107 Microbiology > 310705 Mycology @ 34% 31 BIOLOGICAL SCIENCES > 3105 Genetics > 310507 Genetic immunology @ 33% 31 BIOLOGICAL SCIENCES > 3103 Ecology > 310308 Terrestrial ecology @ 33%
SEO Codes:	28 EXPANDING KNOWLEDGE > 2801 Expanding knowledge > 280102 Expanding knowledge in the biological sciences @ 100%
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