DsRNA-mediated targeting of ribosomal transcripts RPS6 and RPL26 induces long-lasting and significant reductions in fecundity of the vector Aedes aegypti

• Published in: Journal of insect physiology Vol. 90; pp. 17 - 26
• Main Authors: Estep, A.S., Sanscrainte, N.D., Becnel, J.J.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.07.2016
• Subjects: Aedes - genetics; Aedes - physiology; Aedes aegypti; Animals; dsRNA; Fecundity; Female; Fertility; Gene Knockdown Techniques; Gene silencing; Insect Proteins - genetics; Insect Proteins - metabolism; Ixodidae; Oviposition; Ribosomal Proteins - genetics; Ribosomal Proteins - metabolism; Ribosome; RNA, Double-Stranded - genetics; RNA, Double-Stranded - metabolism; siRNA; Trigger; dsRNA; Gene silencing; Ribosome; Trigger; Aedes aegypti; siRNA; Oviposition; Fecundity
• ISSN: 0022-1910; 1879-1611
• DOI: 10.1016/j.jinsphys.2016.05.001

[Display omitted] •DsRNA targeting RPS6 or RPL26 resulted in >88% reduction in fecundity.•RPS6 and RPL26 relative expression was reduced up to 20days post-injection.•DsRNA targeting ribosomal RPL1 or RPS2 reduced expression but not fecundity.•Mutated dsRPS6 indicated complete loss of efficacy with 4 mismatches per siRNA.•Full length dsRPS6 delivered in sugar meals or injected siRPS6 were not effective. Ribosomal transcripts produce critical proteins that are involved in most cellular production processes. Targeting ribosomal transcripts has produced mortality in mites and ticks but the effect of ribosomal transcript knockdown has not been thoroughly examined in mosquitoes. We examine the effects of triggers targeting four ribosomal proteins (RP) transcripts. Although no significant mortality was observed after dsRNA microinjection and subsequent blood feeding, significant contrasts were observed on fecundity. Triggers targeting RPS6 and RPL26 effectively reduced gene expression but more importantly, reduced reproductive output by more than 96% and 91% at the first oviposition while triggers targeting RPL1 and RPS2 did not cause a reduction although gene expression was reduced. Significantly reduced fecundity continued through a second oviposition cycle in dsRPS6 and dsRPL26 cohorts, although the effect was not as strong. Relative gene expression levels confirmed specific transcript knockdown up to 20days post-injection in mosquitoes that did not oviposit or produced reduced clutch sizes. Dissections at 36h post-blood meal indicated defects in oocyte provisioning. The strong phenotype produced by dsRPS6 allowed us to examine the effects in various tissues as well as the dose response, trigger format, delivery method and trigger specificity in Aedes aegypti. Strong knockdown was observed in the abdomen and the ovaries. Greater than 50ng of dsRPS6 significantly reduced fecundity but not when delivered in a sugar meal or as an siRNA. Similar bioassays with mutated dsRPS6 triggers indicates that up to three mismatches per possible siRNA are still effective in reducing fecundity. These studies indicate that while active and effective triggers can be developed for vector species, the lack of an efficient delivery method is the biggest barrier to use as a potential control method.