Robust gut-specific gene expression in transgenic
            <i>Aedes aegypti</i>
            mosquitoes

Moreira, Luciano Andrade; Edwards, Marten J.; Adhami, Faisal; Jasinskiene, Nijole; James, Anthony A.; Jacobs-­Lorena, Marcelo

doi:10.1073/pnas.97.20.10895

Cited by 149 publications

(126 citation statements)

References 18 publications

(22 reference statements)

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“…The temporal expression pattern of AeCPA in midgut epithelial cells appeared to fit the relatively narrow time window described in the scenario above. Moreira et al (18) observed strong marker gene expression under the control of the AeCPA promoter 8-48 h pbm in midguts of A. aegypti. In this study, the temporal expression pattern of the IR construct appeared to coincide with the DENV eclipse phase followed by a slow onset of viral replication within newly infected midgut tissue.…”

Section: Discussionmentioning

confidence: 99%

“…3D). Because the AeCPA promoter activates gene expression in the midgut epithelium (18,19), reduced DENV-2 dissemination in Carb77 could be due to the inability of the virus to reach a concentration threshold in the midgut tissue. Thus, virus amounts escaping the midgut were insufficient to cause infections at later times in salivary glands.…”

Section: Molecular Analysis Of Transgene Expression and Rnaimentioning

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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367

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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Section: Discussionmentioning

confidence: 99%

Section: Molecular Analysis Of Transgene Expression and Rnaimentioning

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

367

375

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“…In this context, we have shown that the Anopheles gambiae and Aedes aegypti carboxypeptidase promoters can be used to drive strong expression of recombinant protein in the midgut of transgenic mosquitoes (5) and also that they can drive the expression of a parasite blocking peptide (6) in transgenic mosquitoes (7). Although the latter results indicate that genetic manipulation of mosquito vectors is a promising strategy for reducing malaria transmission, it is also important to consider that the use of a single effector gene is likely to lead to rapid selection of resistant parasites.…”

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confidence: 99%

Bee Venom Phospholipase Inhibits Malaria Parasite Development in Transgenic Mosquitoes

Moreira

Ito

Ghosh

et al. 2002

Journal of Biological Chemistry

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“…For insects, the most important transposable elements for their transformation success were: Hermes from Musca domestica L. (Warren et al 1994) used to transform A. aegypti (Jasinskiene et al 1998, Kokoza et al 2000, Moreira et al 2000, Pinkerton et al 2000, mariner or Mos1 from Drosophila mauritiana Tsacas & David (Medhora et al 1991) for A. aegypti transformation (Coates et al 1998, Moreira et al 2000; Minos from Drosophila hydei Sturtevant (Franz & Savakis, 1991) used to transform Anopheles stephensi Liston (Catteruccia et al 2000) and piggyBac from Trichoplusia ni Hübner (Cary et al 1989) that has been shown to be very promiscuous in a number of insect orders and also mosquito species [ (Ito et al 2002, Moreira et al 2002, for A. gambiae (Grossman et al 2001) and more recently for Anopheles albimanus Wiedemann (Perera et al 2002)]. …”

Section: Overview Of the Mosquito Transformationmentioning

confidence: 99%

“…Kokoza et al (2000) used the A. aegypti vitellogenin promoter to drive a strong expression of the mosquito defensin (an antibacterial peptide) in the hemolymph of transgenic A. aegypti females. In the mosquito midgut, Moreira et al (2000) demonstrated that both A. gambiae and A. aegypti carboxypeptidases promoters (Edwards et al 1997(Edwards et al , 2000 could drive strong blood-inducible transgene expression in transgenic A. aegypti. Expression was tissue, temporal and sex specific.…”

Section: How To Drive the Expression Of A Transgene?mentioning

confidence: 99%

Transgenic mosquitoes for malaria control: progresses and challenges

Moreira¹,

Jacobs-Lorena²

2003

Neotrop. Entomol.

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Mosquitos Transgênicos Para o Controle da Malária: Progressos e DesafiosRESUMO -A malária mata milhões de pessoas a cada ano e as estratégias atuais de controle da doença, como inseticidas e drogas não têm sido tão eficientes. Por este motivo, novos meios para o combate à malária são de extrema importância. Avanços no estudo do mosquito vetor e sua interação com o parasito da malária fizeram os cientistas pensarem que é possível a manipulação genética dos mosquitos para torná-los vetores ineficientes. Neste artigo, revisamos os avanços na introdução de genes exógenos na linhagem germinativa de mosquitos, a caracterização de promotores específicos de certos tecidos, a identificação de produtos gênicos que bloqueiam o parasita no mosquito, bem como discutimos a recente geração de mosquitos transgênicos, menos eficientes na transmissão de malária. Enquanto muitos progressos foram obtidos, muitos anos de pesquisa são ainda necessários para que mosquitos transgênicos possam ser utilizados na natureza. PALAVRAS-CHAVE: Malária, transgenia, bloqueio da transmissão, PlasmodiumABSTRACT -Malaria kills millions of people every year and the current strategies to control the disease, such as insecticides and drugs have not been completely efficient. Because of that, novel means to fight against malaria are of utmost importance. Advances in the study of the mosquito vector and its interactions with the malaria parasite made scientists think that it is possible to genetically manipulate the mosquitoes to make them inefficient vectors. Here we review the advances on the introduction of foreign genes into the mosquito germ line, the characterization of tissue-specific promoters, the identification of gene products that block development of the parasite in the mosquito, and we discuss the recent generation of transgenic mosquitoes impaired for malaria transmission. While much progress has been made, many years of research are still needed before transgenic mosquitoes can be used in the field.

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Robust gut-specific gene expression in transgenic Aedes aegypti mosquitoes

Cited by 149 publications

References 18 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

Bee Venom Phospholipase Inhibits Malaria Parasite Development in Transgenic Mosquitoes

Transgenic mosquitoes for malaria control: progresses and challenges

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