Vector mosquitoes inflict more human suffering than any other organism—and kill one million people each year. The mosquito genome projects facilitated research in new facets of biology, including functional genetic studies the primary African malaria vector Anopheles gambiae and dengue yellow fever Aedes aegypti. RNA interference- (RNAi-) mediated gene silencing has been used to target genes interest both these disease species. Here, we describe a procedure for preparation...

Despite the devastating impact of mosquito-borne illnesses on human health, surprisingly little is known about mosquito developmental biology, including development olfactory system, a tissue vector importance. Analysis genetics has been hindered by lack means to target specific genes during this sensory system. In investigation, chitosan/siRNA nanoparticles were used semaphorin-1a (sema1a) system in dengue and yellow fever Aedes aegypti. Immunohistochemical analyses anterograde tracing...

The development of sex-specific traits, including the female-specific ability to bite humans and vector disease, is critical for mosquito reproduction pathogen transmission. Doublesex (Dsx), a terminal transcription factor in sex determination pathway, known regulate gene expression during dengue fever Aedes aegypti. Here, effects developmental siRNA-mediated dsx silencing were assessed adult females. Targeting A. aegypti resulted decreased female wing size, correlate body which typically...

Although larviciding can reduce the number of outdoor biting malaria vector mosquitoes, which may help to prevent residual transmission, current larvicide repertoire is faced with great challenges sustainability. The identification new effective, economical, and biorational larvicides could facilitate maintenance expansion practice in integrated mosquito control programmes. Interfering RNA molecules represent a novel class untapped potential for sustainable control. This investigation tested...

New mosquito control strategies are vitally needed to address established arthropod-borne infectious diseases such as dengue and yellow fever emerging Zika chikungunya, all of which transmitted by the disease vector Aedes aegypti. In this investigation, Saccharomyces cerevisiae (baker's yeast) was engineered produce short hairpin RNAs (shRNAs) corresponding aegypti orthologs fasciculation elongation protein zeta 2 (fez2) leukocyte receptor cluster (lrc) member, two genes identified in a...

Vector mosquitoes inflict more human suffering than any other organism—and kill one million people each year. The mosquito genome projects facilitated research in new facets of biology, including functional genetic studies the primary African malaria vector Anopheles gambiae and dengue yellow fever Aedes aegypti. RNA interference- (RNAi-) mediated gene silencing has been used to target genes interest both these disease species. Here, we describe a procedure for preparation...

RNA interference (RNAi), which has facilitated functional characterization of mosquito neural development genes such as the axon guidance regulator semaphorin-1a (sema1a), could one day be applied a new means vector control. Saccharomyces cerevisiae (baker's yeast) may represent an effective interfering expression system that used directly for delivery pesticides to larvae. Here we describe yeast larvicide developed through bioengineering S. express short hairpin (shRNA) targeting conserved...

Background: Drosophila suzukii, or spotted-wing drosophila (SWD), (Diptera: Drosophilidae), are invasive vinegar flies of East Asian origin that have wreaked havoc on the small fruit and berry industry. In locations where SWD well established, weekly chemical insecticide applications necessary, resulting in increased economic costs, unwanted environmental impacts ensuing from loss non-targeted organisms, eventual emergence populations resistant to these insecticides. It is therefore critical...

Essentially nothing is known about the genetic regulation of olfactory system development in vector mosquitoes, which use cues to detect blood meal hosts. Studies Drosophila melanogaster have identified a regulatory matrix transcription factors that controls pupal/adult odorant receptor (OR) gene expression neurons (ORNs). However, it unclear if function D. are required for OR mosquitoes. Furthermore, during larval system, far less complex than pupae/adults, not well understood any insect,...

New mosquito control strategies are vitally needed to address established and emerging arthropod-borne infectious diseases. Here we describe the characterization of a yeast interfering RNA larvicide that was developed through genetic engineering Saccharomyces cerevisiae (baker's yeast) express short hairpin targeting Aedes aegypti synaptotagmin (Aae syt) gene. The effectively silences Aae syt gene, causes defects at larval neural synapse, induces high rates A. mortality in laboratory,...

The existing mosquito pesticide repertoire faces great challenges to sustainability, and new classes of pesticides are vitally needed address established emerging mosquito-borne infectious diseases. RNA interference- (RNAi-) based as a promising biorational control strategy. In this investigation, we describe characterization an interfering (IRP) corresponding the Shaker (Sh) gene, which encodes evolutionarily conserved voltage-gated potassium channel subunit. Delivery IRP Aedes aegypti...

Most animal species exhibit sexually dimorphic behaviors, many of which are linked to reproduction. A number these including blood feeding in female mosquitoes, contribute the global spread vector-borne illnesses. However, knowledge concerning genetic basis traits is limited any organism, especially with respect differences developing nervous system. Custom microarrays were used examine vs. male gene expression pupal head dengue vector mosquito, Aedes aegypti. The spatial patterns a subset...

Despite substantial progress in mosquito genomic and genetic research, few cis-regulatory elements (CREs), DNA sequences that control gene expression, have been identified mosquitoes or other non-model insects. Formaldehyde-assisted isolation of regulatory paired with sequencing, FAIRE-seq, is emerging as a powerful new high-throughput tool for global CRE discovery. FAIRE results the preferential recovery open chromatin fragments are not bound by nucleosomes, an evolutionarily conserved...

Sophisticated tools for manipulation of gene expression in select neurons, including neurons that regulate sexually dimorphic behaviors, are increasingly available analysis genetic model organisms. However, we lack comparable non-model organisms, Aedes aegypti, a vector mosquito which displays behaviors contribute to pathogen transmission. Formaldehyde-assisted isolation regulatory elements followed by sequencing (FAIRE-seq) recently facilitated genome-wide discovery putative A. aegypti...

The global deployment of RNAi yeast insecticides involves transitioning from the use laboratory strains to more robust that are suitable for scaled fermentation. In this investigation, RNA-guided Cas-CLOVER system was used in combination with Piggybac transposase produce Saccharomyces cerevisiae multiple integrated copies Sh.463 short hairpin RNA (shRNA) insecticide expression cassette. This enabled constitutive high-level an insecticidal shRNA corresponding a target sequence is conserved...

Despite the devastating impact of mosquito-borne illnesses on human health, very little is known about mosquito developmental biology, including development visual system. Mosquitoes possess functional adult compound eyes as larvae, a trait that makes them an interesting model in which to study comparative genetics. Here, we functionally characterize system dengue and yellow fever vector Aedes aegypti, use chitosan/siRNA nanoparticles target axon guidance gene semaphorin-1a...

Prevention of mosquito-borne infectious diseases will require new classes environmentally safe insecticides and novel mosquito control technologies. Saccharomyces cerevisiae was engineered to express short hairpin RNA (shRNA) corresponding Rbfox1 genes. The yeast induced target gene silencing, resulting in larval death that observed both laboratory outdoor semi-field trials conducted on Aedes aegypti. High levels mortality were also during simulated field which adult females consumed...

Abstract Aedes mosquitoes (Diptera: Culicidae), principle vectors of several arboviruses, typically lay eggs in man‐made water‐filled containers located near human dwellings. Given the widespread emergence insecticide resistance, stable and biofriendly alternatives for mosquito larviciding are needed. Laboratory studies have demonstrated that inactivated yeast interfering RNA tablets targeting key larval developmental genes can be used to facilitate effective larvicidal activity while also...

Concerns for widespread insecticide resistance and the unintended impacts of insecticides on nontarget organisms have generated a pressing need mosquito control innovations. A yeast RNAi-based that targets conserved site in Irx family genes, but which has not yet been identified genomes organisms, was developed characterized. Saccharomyces cerevisiae constructed to express short hairpin RNA (shRNA) matching target induced significant Aedes aegypti larval death both lab trials outdoor...

Abstract Although many putative long non-coding RNA (lncRNA) genes have been identified in insect genomes, few of these functionally validated. A screen for female-specific larvicides that facilitate Aedes aegypti male sex separation uncovered multiple interfering RNAs with target sites lncRNA located the M/m locus region, including loci within or tightly linked to determination locus. Larval consumption a Saccharomyces cerevisiae (yeast) strain engineered express corresponding transcripts...