This site needs JavaScript to work properly. Please enable it to take advantage of the complete set of features!
Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log in
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
doi: 10.1186/1475-2875年12月14日2.

Development of a population suppression strain of the human malaria vector mosquito, Anopheles stephensi

Affiliations

Development of a population suppression strain of the human malaria vector mosquito, Anopheles stephensi

Osvaldo Marinotti et al. Malar J. .

Abstract

Background: Transgenic mosquito strains are being developed to contribute to the control of dengue and malaria transmission. One approach uses genetic manipulation to confer conditional, female-specific dominant lethality phenotypes. Engineering of a female-specific flightless phenotype provides a sexing mechanism essential for male-only mosquito, release approaches that result in population suppression of target vector species.

Methods: An approach that uses a female-specific gene promoter and antibiotic-repressible lethal factor to produce a sex-specific flightless phenotype was adapted to the human malaria vector, Anopheles stephensi. Transposon- and site-specific recombination-mediated technologies were used to generate a number of transgenic An. stephensi lines that when combined through mating produced the phenotype of flight-inhibited females and flight-capable males.

Results: The data shown here demonstrate the successful engineering of a female-specific flightless phenotype in a malaria vector. The flightless phenotype was repressible by the addition of tetracycline to the larval diet. This conditional phenotype allows the rearing of the strains under routine laboratory conditions. The minimal level of tetracycline that rescues the flightless phenotype is higher than that found as an environmental contaminant in circumstances where there is intensive use of antibiotics.

Conclusions: These studies support the further development of flightless female technology for applications in malaria control programmes that target the vectors.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Schematic representations of the transformation constructs OX3545, AsOX3545 and OX3547, and the mechanism of action of the double transgene-based female-specific RIDL. OX3545 and OX3547 were previously described in detail [2]. AsOX3545 is a similar to OX3545, with the Anopheles stephensi Actin-4 regulatory sequences replacing the Aedes aegypti Actin-4 sequence. Actin 4 promoters (Act4P) drive the expression of the tetracycline-repressible transactivator protein (tTa) in the flight muscles of adult female mosquitoes. tTA binds tetO in the absence of tetracycline, driving expression of the effector molecule, Nuclear Inhibitor of PP1 (Nipp1Dm) [22,23], leading to disruption of normal cell functions. Tetracycline prevents tTa binding to tetO abolishing expression of Nipp1Dm. The positions of the SpeI recognition and cleavage sites in OX3545, AsOX3545 and OX3547 are indicated by vertically oriented arrows. Horizontally oriented arrows indicate the direction of transcription for each component of the constructs. Abbreviations: pBac R and L, piggyBac right and left inverted repeats, respectively; attB, φC31 recombination site; DsRed2, red fluorescent protein open reading frame; IE1, baculovirus immediate-early gene promoter driving DsRed2 expressed throughout the body; ×ばつP3, artificial promoter consisting of a multimer of the binding site (P3), driving high levels of expression in the eye.
Figure 2
Figure 2
Southern blot detection of transgenes integrated into the genomes of Anopheles stephensi driver and effector transgenic lines. Genomic DNA extracted from a pool of mosquitoes from each line was digested with SpeI, fractionated by agarose gel electrophoresis, transferred to a Zeta-probe GT membrane and hybridized to a 32P-labelled DsRed2 open-reading frame probe. DAa, DAs and E and the numbers on top of the figures indicate the individual transgenic lines originated from injection of the OX3545 and AsOX3534 driver and OX3547 effector constructs, respectively.

References

    1. Thomas DD, Donnelly CA, Wood RJ, Alphey LS. Insect population control using a dominant, repressible, lethal genetic system. Science. 2010;287:2474–2476. - PubMed
    1. Fu G, Lees RS, Nimmo D, Aw D, Jin L, Gray P, Berendonk TU, White-Cooper H, Scaife S, Kim Phuc H, Marinotti O, Jasinskiene N, James AA, Alphey L. Female-specific flightless phenotype for mosquito control. Proc Natl Acad Sci U S A. 2010;107:4550–4554. doi: 10.1073/pnas.1000251107. - DOI - PMC - PubMed
    1. Black WC 4th, Alphey L, James AA. Why RIDL is not SIT. Trends Parasitol. 2011;27:362–370. doi: 10.1016/j.pt.201104004. - DOI - PubMed
    1. Labbé GM, Scaife S, Morgan SA, Curtis ZH, Alphey L. Female-specific flightless (fsRIDL) phenotype for control of Aedes albopictus. PLoS Negl Trop Dis. 2012;6:e1724. doi: 10.1371/journal.pntd.0001724. - DOI - PMC - PubMed
    1. Alphey L, Benedict M, Bellini R, Clark GG, Dame DA, Service MW, Dobson SL. Sterile-insect methods for control of mosquito-borne diseases: an analysis. Vector Borne Zoonotic Dis. 2010;10:295–311. doi: 10.1089/vbz.2009.0014. - DOI - PMC - PubMed

Publication types

Substances

Cite

AltStyle によって変換されたページ (->オリジナル) /