Stable transformation of the yellow fever mosquito,
            <i>Aedes aegypti</i>
            , with the
            <i>Hermes</i>
            element from the housefly

Jasinskiene, Nijole; Coates, Craig J.; Benedict, Mark Q.; Cornel, Anthony J.; Rafferty, Cristina Salazar; James, Anthony A.; Collins, Frank H.

doi:10.1073/pnas.95.7.3743

Cited by 332 publications

(221 citation statements)

References 19 publications

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“…Germ-line transformations were carried out as described (34,36). Each surviving G 0 male was outcrossed to 20 HWE females.…”

Section: Germ-line Transformation Of a Aegypti And Establishment Of mentioning

confidence: 99%

“…DNA extractions and Southern blot analyses followed the procedures described (34,36). About 20 g of extracted total DNA was digested either with EcoRI, KpnI, or PstI and column-purified (QIAquick PCR Purification Kit, Qiagen, Valencia, CA) followed by electrophoresis in a 0.8% agarose gel and blotting onto a positively charged nylon membrane (Ambion, Austin, TX).…”

Section: Germ-line Transformation Of a Aegypti And Establishment Of mentioning

confidence: 99%

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Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti

Franz

Sánchez-Vargas

Adelman

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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368

375

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Mosquitoes (Aedes aegypti) were genetically modified to exhibit impaired vector competence for dengue type 2 viruses (DENV-2). We exploited the natural antiviral RNA interference (RNAi) pathway in the mosquito midgut by constructing an effector gene that expresses an inverted-repeat (IR) RNA derived from the premembrane protein coding region of the DENV-2 RNA genome. The A. aegypti carboxypeptidase A promoter was used to express the IR RNA in midgut epithelial cells after ingestion of a bloodmeal. The promoter and effector gene were inserted into the genome of a white-eye Puerto Rico Rexville D (Higgs' white eye) strain by using the nonautonomous mariner MosI transformation system. A transgenic family, Carb77, expressed IR RNA in the midgut after a bloodmeal. Carb77 mosquitoes ingesting an artificial bloodmeal containing DENV-2 exhibited marked reduction of viral envelope antigen in midguts and salivary glands after infection. DENV-2 titration of individual mosquitoes showed that most Carb77 mosquitoes poorly supported virus replication. Transmission in vitro of virus from the Carb77 line was significantly diminished when compared to control mosquitoes. The presence of DENV-2-derived siRNAs in RNA extracts from midguts of Carb77 and the loss of the resistance phenotype when the RNAi pathway was interrupted proved that DENV-2 resistance was caused by a RNAi response. Engineering of transgenic A. aegypti that show a high level of resistance against DENV-2 provides a powerful tool for developing population replacement strategies to control transmission of dengue viruses.RNA silencing ͉ transgenesis ͉ genetic control ͉ mosquito ͉ dengue disease D engue viruses (DENV) [Flaviviridae; Flavivirus; DENV types 1-4 (DENV-1-4)] threaten public health in Ͼ100 countries and infect an estimated 50 million people annually (1, 2). The mosquito, Aedes aegypti, is the principal vector for epidemic dengue disease (3). The urban DENV transmission cycle involves only humans and mosquitoes. No DENV vaccines are currently available, and vector control strategies that minimize human-mosquito contact have largely failed (4, 5). New control strategies are needed. One possible strategy is to replace vector populations competent to transmit DENVs with pathogenincompetent vectors (6). The essential features of this genetic control strategy are to identify genes that express antiviral molecules in the vector, link this gene (or genes) to a genetic drive system [transposable elements (TE), meiotic drive, or homing endonuclease genes] and introgress the gene(s) into field populations (7-9). A key step in developing this control strategy is to identify effector genes that, when expressed in the vector, inhibit DENV replication. Proof of principle for RNA interference (RNAi)-like disruption of DENV-2 vector competence was previously demonstrated by using a nonheritable alphavirus expression system (10). Applying the principle of heritable gene silencing in transgenic Drosophila melanogaster, Caenorhabditis elegans, and A. aegypti (11-13), we gene...

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“…Germ-line transformations were carried out as described (34,36). Each surviving G 0 male was outcrossed to 20 HWE females.…”

Section: Germ-line Transformation Of a Aegypti And Establishment Of mentioning

confidence: 99%

Section: Germ-line Transformation Of a Aegypti And Establishment Of mentioning

confidence: 99%

Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti

Franz

Sánchez-Vargas

Adelman

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

368

375

View full text Add to dashboard Cite

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“…Although P has transferred horizontally among Drosophila species, it is considered a ''dead'' element in other insects (26). Thus, the choice of TE for donating the terminal inverted repeats and transposase for the gene drive systems here was limited to those demonstrated to mobilize in A. aegypti: MosI (7), piggyBac (27), and Hermes (8). Although our data demonstrate that noscontrolled MosI can mobilize a plasmid-based element, remobilization, movement of a chromosomally integrated transgene, has yet to be observed.…”

Section: Discussionmentioning

confidence: 99%

“…These strategies benefit from the availability of transgenesis technologies, and stable integration of engineered genes into A. aegypti, using transposable elements (TEs) (7,8) is now routine (9). Population replacement strategies also depend on the development of effector genes to interfere with and prevent pathogen transmission.…”

Section: Dengue ͉ Gene Drive ͉ Transposonmentioning

confidence: 99%

nanos gene control DNA mediates developmentally regulated transposition in the yellow fever mosquito Aedes aegypti

Adelman

Jasinskiene²,

Onal³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Transposable elements (TEs) are proposed as a basis for developing drive systems to spread pathogen resistance genes through vector mosquito populations. The use of transcriptional and translational control DNA elements from genes expressed specifically in the insect germ line to mediate transposition offers possibilities for mitigating some of the concerns about transgene behavior in the target vector species and eliminating effects on nontarget organisms. Here, we describe the successful use of the promoter and untranslated regions from the nanos (nos) orthologous gene of the yellow fever mosquito, Aedes aegypti, to control sex-and tissuespecific expression of exogenously derived mariner MosI transposase-encoding DNA. Transgenic mosquitoes expressed transposase mRNA in abundance near or equal to the endogenous nos transcript and exclusively in the female germ cells. In addition, MosI mRNA was deposited in developing oocytes and localized and maintained at the posterior pole during early embryonic development. Importantly, four of five transgenic lines examined were capable of mobilizing a second MosI transgene into the mosquito genome, indicating that functional transposase was being produced. Thus, the nos control sequences show promise as part of a TE-based gene drive system. dengue ͉ gene drive ͉ transposon

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“…Recent efforts to genetically manipulate mosquitoes to control mosquito-transmitted diseases have raised considerable interest in the KMO gene, because it can serve as an excellent marker that indicates the successful production of transgenic Ae. aegypti Jasinskiene et al, 1998;Sethuraman and O'Brochta, 2005). Sequence analysis of the wild-type and mutant kh w cDNAs revealed a deletion of 162 nucleotides in the mutant Ae.…”

Section: Kynurenine 3-monooxygenase (Kmo)mentioning

confidence: 99%

The tryptophan oxidation pathway in mosquitoes with emphasis on xanthurenic acid biosynthesis

Han¹,

Beerntsen²,

Li³

2007

Journal of Insect Physiology

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Oxidation of tryptophan to kynurenine and 3-hydroxykynurenine (3-HK) is the major catabolic pathway in mosquitoes. However, 3-HK is oxidized easily under physiological conditions, resulting in the production of reactive radical species. To overcome this problem, mosquitoes have developed an efficient mechanism to prevent 3-HK from accumulating by converting this chemically reactive compound to the chemically stable xanthurenic acid. Interestingly, 3-HK is a precursor for the production of compound eye pigments during the pupal and early adult stages; consequently, mosquitoes need to preserve and transport 3-HK for compound eye pigmentation in pupae and adults. This review summarizes the tryptophan oxidation pathway, compares and contrasts the mosquito tryptophan oxidation pathway with other model species, and discusses possible driving forces leading to the functional adaptation and evolution of enzymes involved in the mosquito tryptophan oxidation pathway.

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Stable transformation of the yellow fever mosquito, Aedes aegypti , with the Hermes element from the housefly

Cited by 332 publications

References 19 publications

Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti

Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti

nanos gene control DNA mediates developmentally regulated transposition in the yellow fever mosquito Aedes aegypti

The tryptophan oxidation pathway in mosquitoes with emphasis on xanthurenic acid biosynthesis

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