RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari
Abstract RNA interference (RNAi) is an effective approach to suppress gene expression and monitor gene regulation. Despite its wide application, its use is limited in certain taxonomic groups, including cnidarians. Myxozoans are a unique group of cnidarian parasites that diverged from their free-liv...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-02-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-54171-0 |
| _version_ | 1797274616144592896 |
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| author | Jiří Kyslík Ana Born-Torrijos Astrid S. Holzer Anush Kosakyan |
| author_facet | Jiří Kyslík Ana Born-Torrijos Astrid S. Holzer Anush Kosakyan |
| author_sort | Jiří Kyslík |
| collection | DOAJ |
| description | Abstract RNA interference (RNAi) is an effective approach to suppress gene expression and monitor gene regulation. Despite its wide application, its use is limited in certain taxonomic groups, including cnidarians. Myxozoans are a unique group of cnidarian parasites that diverged from their free-living ancestors about 600 million years ago, with several species causing acute disease in farmed and wild fish populations. In this pioneering study we successfully applied RNAi in blood stages of the myxozoan Sphaerospora molnari, combining a dsRNA soaking approach, real-time PCR, confocal microscopy, and Western blotting. For proof of concept, we knocked down two unusual actins, one of which is known to play a critical role in S. molnari cell motility. We observed intracellular uptake of dsRNA after 30 min and accumulation in all cells of the typical myxozoan cell-in-cell structure. We successfully knocked down actin in S. molnari in vitro, with transient inhibition for 48 h. We observed the disruption of the cytoskeletal network within the primary cell and loss of the characteristic rotational cell motility. This RNAi workflow could significantly advance functional research within the Myxozoa, offering new prospects for investigating therapeutic targets and facilitating drug discovery against economically important fish parasites. |
| first_indexed | 2024-03-07T15:00:50Z |
| format | Article |
| id | doaj.art-d5b5d675e553419b9e68d1ca596c6f2f |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-07T15:00:50Z |
| publishDate | 2024-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-d5b5d675e553419b9e68d1ca596c6f2f2024-03-05T19:09:58ZengNature PortfolioScientific Reports2045-23222024-02-0114111510.1038/s41598-024-54171-0RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnariJiří Kyslík0Ana Born-Torrijos1Astrid S. Holzer2Anush Kosakyan3Institute of Parasitology, Biology Centre, Academy of Sciences of the Czech RepublicInstitute of Parasitology, Biology Centre, Academy of Sciences of the Czech RepublicInstitute of Parasitology, Biology Centre, Academy of Sciences of the Czech RepublicInstitute of Parasitology, Biology Centre, Academy of Sciences of the Czech RepublicAbstract RNA interference (RNAi) is an effective approach to suppress gene expression and monitor gene regulation. Despite its wide application, its use is limited in certain taxonomic groups, including cnidarians. Myxozoans are a unique group of cnidarian parasites that diverged from their free-living ancestors about 600 million years ago, with several species causing acute disease in farmed and wild fish populations. In this pioneering study we successfully applied RNAi in blood stages of the myxozoan Sphaerospora molnari, combining a dsRNA soaking approach, real-time PCR, confocal microscopy, and Western blotting. For proof of concept, we knocked down two unusual actins, one of which is known to play a critical role in S. molnari cell motility. We observed intracellular uptake of dsRNA after 30 min and accumulation in all cells of the typical myxozoan cell-in-cell structure. We successfully knocked down actin in S. molnari in vitro, with transient inhibition for 48 h. We observed the disruption of the cytoskeletal network within the primary cell and loss of the characteristic rotational cell motility. This RNAi workflow could significantly advance functional research within the Myxozoa, offering new prospects for investigating therapeutic targets and facilitating drug discovery against economically important fish parasites.https://doi.org/10.1038/s41598-024-54171-0 |
| spellingShingle | Jiří Kyslík Ana Born-Torrijos Astrid S. Holzer Anush Kosakyan RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari Scientific Reports |
| title | RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari |
| title_full | RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari |
| title_fullStr | RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari |
| title_full_unstemmed | RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari |
| title_short | RNAi-directed knockdown in the cnidarian fish blood parasite Sphaerospora molnari |
| title_sort | rnai directed knockdown in the cnidarian fish blood parasite sphaerospora molnari |
| url | https://doi.org/10.1038/s41598-024-54171-0 |
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