Dataset Information

RNAi by Soaking Aedes aegypti Pupae in dsRNA.

ABSTRACT: RNA-interference (RNAi) is a standard technique for functional genomics in adult mosquitoes. However, RNAi in immature, aquatic mosquito stages has been challenging. Several studies have shown successful larval RNAi, usually in combination with a carrier molecule. Except for one study in malaria mosquito, Anopheles gambiae, none of the previous studies has explored RNAi in mosquito pupae. Even in the study that used RNAi in pupae, double stranded RNA (dsRNA) was introduced by microinjection. Here, we describe a successful method by soaking pupae in water containing dsRNA without any carrier or osmotic challenge. The knockdown persisted into adulthood. We expect that this simple procedure will be useful in the functional analysis of genes that highly express in pupae or newly emerged adults.

SUBMITTER: Arshad F

PROVIDER: S-EPMC8307132 | biostudies-literature | 2021 Jul

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

RNAi by Soaking <i>Aedes aegypti</i> Pupae in dsRNA.

Arshad Fiza F Sharma Arvind A Lu Charleen C Gulia-Nuss Monika M

Insects 20210713 7

RNA-interference (RNAi) is a standard technique for functional genomics in adult mosquitoes. However, RNAi in immature, aquatic mosquito stages has been challenging. Several studies have shown successful larval RNAi, usually in combination with a carrier molecule. Except for one study in malaria mosquito, <i>Anopheles gambiae</i>, none of the previous studies has explored RNAi in mosquito pupae. Even in the study that used RNAi in pupae, double stranded RNA (dsRNA) was introduced by microinjecti ...[more]

PMID: 34357293

Similar Datasets

Project description:BackgroundThe yellow fever mosquito, Aedes aegypti, vectors several pathogens responsible for human diseases. As a result, this mosquito species is a priority for control by mosquito control districts in Florida. With insecticide resistance development becoming a concern, alternative control strategies are needed for Ae. aegypti. Sterile insect technique (SIT) is an increasingly popular option that is being explored as a practical area-wide control method. However, questions about sterile male performance persist. The objectives of this study were to determine the extent to which hypoxia exposure prior to and during irradiation effects the longevity, activity and mating competitiveness of sterile male Ae. aegypti.MethodsMale longevity was monitored and analyzed using Cox regression. Mosquito activity was recorded by an infrared beam sensor rig that detected movement. Competing models were created to analyze movement data. Fecundity and fertility were measured in females mated with individual males by treatment and analyzed using one-way ANOVAs. Mating competition studies were performed to compare both hypoxia and normoxia treated sterile males to fertile males. Competitiveness of groups was compared using Fried's competitiveness index.ResultsFirst, we found that subjecting Ae. aegypti pupae to 1 h of severe hypoxia (< 1 kPa O2) did not directly increase mortality. One hour of hypoxia was found to prevent decreases in longevity of irradiated males compared to males irradiated in normoxic conditions. Exposure to hypoxia prior to irradiation did not significantly improve activity of sterile males except at the highest doses of radiation. Hypoxia did significantly increase the required dose of radiation to achieve > 95% male sterility compared to males irradiated under normoxic conditions. Males sterilized after an hour in hypoxic conditions were significantly more competitive against fertile males compared to males irradiated under normoxic conditions despite requiring a higher dose of radiation to achieve sterility.ConclusionsHypoxia was found to greatly improve key performance metrics in sterile male Ae. aegypti without any significant drawbacks. Little work other than increasing the target dose for sterility needs to be conducted to incorporate hypoxia into SIT programs. These results suggest that SIT programs should consider including hypoxia in their sterile male production workflow.

Dataset Information

RNAi by Soaking Aedes aegypti Pupae in dsRNA.

Publications

RNAi by Soaking <i>Aedes aegypti</i> Pupae in dsRNA.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets