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To manipulate and decipher gene function, rationally designed small interfering RNA (siRNA) can initiate the destruction of homologous messenger RNA (mRNA), resulting in the \u201cknockdown\u201d of target gene expression. Here, we investigate whether siRNA can be used to manipulate gene expression in <jats:italic>B. dendrobatidis<\/jats:italic> 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\u201348\u2009h after delivery of siRNA targeting glutamate\u2013cysteine ligase, with a maximum of ~56% reduction in target transcripts occurring at 36\u2009h. A second siRNA design targeting glutamate\u2010cysteine 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 <jats:italic>B. dendrobatidis<\/jats:italic>, 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 <jats:italic>B. dendrobatidis.<\/jats:italic><\/jats:p>","DOI":"10.1002\/jobm.202400081","type":"journal-article","created":{"date-parts":[[2024,6,21]],"date-time":"2024-06-21T07:25:12Z","timestamp":1718954712000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Small Interfering RNA Mediated Messenger RNA Knockdown in the Amphibian Pathogen <i>Batrachochytrium dendrobatidis<\/i>"],"prefix":"10.1002","volume":"64","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2539-2755","authenticated-orcid":false,"given":"Rebecca J.","family":"Webb","sequence":"first","affiliation":[{"name":"One Health Research Group, Melbourne Veterinary School University of Melbourne Werribee Victoria Australia"},{"name":"Australian Institute of Tropical Health and Medicine James Cook Univiersity Townsville Queensland Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexandra A.","family":"Roberts","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine James Cook Univiersity Townsville Queensland Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Catherine","family":"Rush","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine James Cook Univiersity Townsville Queensland Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lee F.","family":"Skerratt","sequence":"additional","affiliation":[{"name":"One Health Research Group, Melbourne Veterinary School University of Melbourne Werribee Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark L.","family":"Tizard","sequence":"additional","affiliation":[{"name":"Australian Centre for Disease Preparedness CSIRO Health and Biosecurity Geelong Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lee","family":"Berger","sequence":"additional","affiliation":[{"name":"One Health Research Group, Melbourne Veterinary School University of Melbourne Werribee Victoria Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,6,21]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/418244a"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/1755-0998.12359"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature01278"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1159\/000078214"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.106.084517"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-41832-8"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1359"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fsi.2012.11.037"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1001160"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1093\/mmy\/myt006"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005901"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-25434-4"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80620-0"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/35053110"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1064023"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/35005107"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1102513"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(01)00581-5"},{"key":"e_1_2_10_20_1","first-page":"36","article-title":"Genetic Engineering of Fungal Cells","volume":"3","author":"Moore M. 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