2024Journal of Medical EntomologyOpen access

Efficient RNAi knockdown at 20 °C in Aedes albopictus (Diptera: Culicidae)

Mara Heilig, Peter Armbruster

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Abstract

RNA interference (RNAi) is a conserved cellular pathway found in nearly all eukaryotes that leads to the silencing of target mRNAs. Using RNAi as a mechanism to knockdown specific genes has enabled functional and reverse genetics studies in a wide range of eukaryotes. Previous work suggests that RNAi is inhibited at lower temperatures, potentially limiting the possibility to perform knockdown studies on ecologically relevant phenotypes that are only expressed at low temperatures. To determine whether RNAi is inhibited at low temperatures in Aedes albopictus (Skuse), we injected mosquitoes reared at 20 ± 1 °C, with dsRNA targeting yellow-g2 and compared knockdown efficacy to mosquitoes injected at 26.5 ± 1 °C. Our results demonstrate efficient knockdown at both temperatures, thereby establishing the feasibility of RNAi for functional genetic studies in A. albopictus at low temperatures.

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RNA interference (RNAi) is a conserved cellular pathway found in nearly all eukaryotes that leads to the silencing of target mRNAs. Using RNAi as a mechanism to knockdown specific genes has enabled functional and reverse genetics studies in a wide range of eukaryotes. Previous work suggests that RNAi is inhibited at lower temperatures, potentially limiting the possibility to perform knockdown studies on ecologically relevant phenotypes that are only expressed at low temperatures. To determine whether RNAi is inhibited at low temperatures in Aedes albopictus (Skuse), we injected mosquitoes reared at 20 ± 1 °C, with dsRNA targeting yellow-g2 and compared knockdown efficacy to mosquitoes injected at 26.5 ± 1 °C. Our results demonstrate efficient knockdown at both temperatures, thereby establishing the feasibility of RNAi for functional genetic studies in A. albopictus at low temperatures.

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Available abstract

RNA interference (RNAi) is a conserved cellular pathway found in nearly all eukaryotes that leads to the silencing of target mRNAs. Using RNAi as a mechanism to knockdown specific genes has enabled functional and reverse genetics studies in a wide range of eukaryotes. Previous work suggests that RNAi is inhibited at lower temperatures, potentially limiting the possibility to perform knockdown studies on ecologically relevant phenotypes that are only expressed at low temperatures. To determine whether RNAi is inhibited at low temperatures in Aedes albopictus (Skuse), we injected mosquitoes reared at 20 ± 1 °C, with dsRNA targeting yellow-g2 and compared knockdown efficacy to mosquitoes injected at 26.5 ± 1 °C. Our results demonstrate efficient knockdown at both temperatures, thereby establishing the feasibility of RNAi for functional genetic studies in A. albopictus at low temperatures.

Key concepts: Gene knockdown, RNA interference, Biology, Aedes albopictus, Gene silencing, RNA silencing, Knockdown resistance, Small interfering RNA

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Efficient RNAi knockdown at 20 °C in Aedes albopictus (Diptera: Culicidae) — Research Paper | ScholarLens