RNAi-mediated knockdown of daf-12 in the model parasitic nematode Strongyloides ratti.
The gene daf-12 has long shown to be involved in the dauer pathway in Caenorhabditis elegans (C. elegans). Due to the similarities of the dauer larvae of C. elegans and infective larvae of certain parasitic nematodes such as Strongyloides spp., this gene has also been suspected to be involved in the...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2019-03-01
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| Series: | PLoS Pathogens |
| Online Access: | http://europepmc.org/articles/PMC6457571?pdf=render |
| _version_ | 1819128189291593728 |
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| author | Alex Dulovic Adrian Streit |
| author_facet | Alex Dulovic Adrian Streit |
| author_sort | Alex Dulovic |
| collection | DOAJ |
| description | The gene daf-12 has long shown to be involved in the dauer pathway in Caenorhabditis elegans (C. elegans). Due to the similarities of the dauer larvae of C. elegans and infective larvae of certain parasitic nematodes such as Strongyloides spp., this gene has also been suspected to be involved in the development of infective larvae. Previous research has shown that the application of dafachronic acid, the steroid hormone ligand of DAF-12 in C. elegans, affects the development of infective larvae and metabolism in Strongyloides. However, a lack of tools for either forward or reverse genetics within Strongyloides has limited studies of gene function within these important parasites. After determining whether Strongyloides had the requisite proteins for RNAi, we developed and report here the first successful RNAi by soaking protocol for Strongyloides ratti (S. ratti) and use this protocol to study the functions of daf-12 within S. ratti. Suppression of daf-12 in S. ratti severely impairs the formation of infective larvae of the direct cycle and redirects development towards the non-infective (non-dauer) free-living life cycle. Further, daf-12(RNAi) S. ratti produce slightly but significantly fewer offspring and these offspring are developmentally delayed or incapable of completing their development to infective larvae (L3i). Whilst the successful daf-12(RNAi) L3i are still able to infect a new host, the resulting infection is less productive and shorter lived. Further, daf-12 knockdown affects metabolism in S. ratti resulting in a shift from aerobic towards anaerobic fat metabolism. Finally, daf-12(RNAi) S. ratti have reduced tolerance of temperature stress. |
| first_indexed | 2024-12-22T08:23:52Z |
| format | Article |
| id | doaj.art-39f9a81debaf4597ade89903f1e723a2 |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-22T08:23:52Z |
| publishDate | 2019-03-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-39f9a81debaf4597ade89903f1e723a22022-12-21T18:32:40ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742019-03-01153e100770510.1371/journal.ppat.1007705RNAi-mediated knockdown of daf-12 in the model parasitic nematode Strongyloides ratti.Alex DulovicAdrian StreitThe gene daf-12 has long shown to be involved in the dauer pathway in Caenorhabditis elegans (C. elegans). Due to the similarities of the dauer larvae of C. elegans and infective larvae of certain parasitic nematodes such as Strongyloides spp., this gene has also been suspected to be involved in the development of infective larvae. Previous research has shown that the application of dafachronic acid, the steroid hormone ligand of DAF-12 in C. elegans, affects the development of infective larvae and metabolism in Strongyloides. However, a lack of tools for either forward or reverse genetics within Strongyloides has limited studies of gene function within these important parasites. After determining whether Strongyloides had the requisite proteins for RNAi, we developed and report here the first successful RNAi by soaking protocol for Strongyloides ratti (S. ratti) and use this protocol to study the functions of daf-12 within S. ratti. Suppression of daf-12 in S. ratti severely impairs the formation of infective larvae of the direct cycle and redirects development towards the non-infective (non-dauer) free-living life cycle. Further, daf-12(RNAi) S. ratti produce slightly but significantly fewer offspring and these offspring are developmentally delayed or incapable of completing their development to infective larvae (L3i). Whilst the successful daf-12(RNAi) L3i are still able to infect a new host, the resulting infection is less productive and shorter lived. Further, daf-12 knockdown affects metabolism in S. ratti resulting in a shift from aerobic towards anaerobic fat metabolism. Finally, daf-12(RNAi) S. ratti have reduced tolerance of temperature stress.http://europepmc.org/articles/PMC6457571?pdf=render |
| spellingShingle | Alex Dulovic Adrian Streit RNAi-mediated knockdown of daf-12 in the model parasitic nematode Strongyloides ratti. PLoS Pathogens |
| title | RNAi-mediated knockdown of daf-12 in the model parasitic nematode Strongyloides ratti. |
| title_full | RNAi-mediated knockdown of daf-12 in the model parasitic nematode Strongyloides ratti. |
| title_fullStr | RNAi-mediated knockdown of daf-12 in the model parasitic nematode Strongyloides ratti. |
| title_full_unstemmed | RNAi-mediated knockdown of daf-12 in the model parasitic nematode Strongyloides ratti. |
| title_short | RNAi-mediated knockdown of daf-12 in the model parasitic nematode Strongyloides ratti. |
| title_sort | rnai mediated knockdown of daf 12 in the model parasitic nematode strongyloides ratti |
| url | http://europepmc.org/articles/PMC6457571?pdf=render |
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