Systemic gene silencing from oral uptake of dsRNA in Litopenaeus vannamei requires both clathrin-mediated endocytosis and LvSID-1
RNA interference (RNAi) is a powerful technique that has been widely applied to inhibit both DNA and RNA viruses in shrimp. To accomplish RNAi in shrimp, dsRNA is often delivered by direct injection or feeding, although the latter method has a much lower efficacy. Therefore, understanding the molecu...
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| Main Authors: | , , |
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| Format: | Article |
| Published: |
2022
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| Online Access: | https://repository.li.mahidol.ac.th/handle/123456789/73079 |
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| Institution: | Mahidol University |
| Summary: | RNA interference (RNAi) is a powerful technique that has been widely applied to inhibit both DNA and RNA viruses in shrimp. To accomplish RNAi in shrimp, dsRNA is often delivered by direct injection or feeding, although the latter method has a much lower efficacy. Therefore, understanding the molecular mechanisms underlying the cellular uptake of dsRNA after oral feeding may help in improving the potency of fed RNAi in shrimp. Chlorpromazine was used to block clathrin-mediated endocytosis before feeding a formulated food containing dsRNA specific to shrimp the Rab7 transcript for 6 days. Suppression of Rab7 expression was seen only in the hepatopancreas and gill tissues of the shrimp fed without drug, and suppression was lost when shrimp were pre-treated with chlorpromazine. The results indicate that clathrin-mediated endocytosis participates in dsRNA uptake from the diet into shrimp hepatopancreas. Up-regulation of LvSID-1 was also observed in the gill of Rab7-silenced shrimp fed with the dsRab7 diet, indicating that the RNAi signal derived from feeding was systemically spread by the hemolymph, possibly in the form of long dsRNA. Moreover, induction of LvSID-1 expression before ingestion of dsSTAT improved suppression levels of STAT mRNA in gill tissues, showing that the fed RNAi signal was taken up into gill cells via the LvSID-1 protein. This study reports for the first time on the involvement of both clathrin-mediated endocytosis and LvSID-1 in systemic gene silencing after oral uptake of dsRNA in shrimp, which could help in improving the potency of antiviral strategies in shrimp by fed with RNAi in the future. |
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