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01.12.2012 | Methodology | Ausgabe 1/2012 Open Access

Malaria Journal 1/2012

High-throughput sorting of mosquito larvae for laboratory studies and for future vector control interventions

Zeitschrift:
Malaria Journal > Ausgabe 1/2012
Autoren:
Eric Marois, Christina Scali, Julien Soichot, Christine Kappler, Elena A Levashina, Flaminia Catteruccia
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1475-2875-11-302) contains supplementary material, which is available to authorized users.
Elena A Levashina and Flaminia Catteruccia contributed equally to this work.

Competing interests

The authors declare that they have no competing interests.

Authors’ contributions

CS and FC designed and constructed the DSX plasmid and the transgenic mosquito strain. EM designed methods, performed experiments and drafted the manuscript. JS and CK generated and mapped the FK X line, respectively. EAL and FC conceived the study, participated to its design and co-ordination and drafted the manuscript. All authors read and approved the final manuscript.

Abstract

Background

Mosquito transgenesis offers new promises for the genetic control of vector-borne infectious diseases such as malaria and dengue fever. Genetic control strategies require the release of large number of male mosquitoes into field populations, whether they are based on the use of sterile males (sterile insect technique, SIT) or on introducing genetic traits conferring refractoriness to disease transmission (population replacement). However, the current absence of high-throughput techniques for sorting different mosquito populations impairs the application of these control measures.

Methods

A method was developed to generate large mosquito populations of the desired sex and genotype. This method combines flow cytometry and the use of Anopheles gambiae transgenic lines that differentially express fluorescent markers in males and females.

Results

Fluorescence-assisted sorting allowed single-step isolation of homozygous transgenic mosquitoes from a mixed population. This method was also used to select wild-type males only with high efficiency and accuracy, a highly desirable tool for genetic control strategies where the release of transgenic individuals may be problematic. Importantly, sorted males showed normal mating ability compared to their unsorted brothers.

Conclusions

The developed method will greatly facilitate both laboratory studies of mosquito vectorial capacity requiring high-throughput approaches and future field interventions in the fight against infectious disease vectors.
Zusatzmaterial
Additional file 1: Description of the FK X transgenic construct.(DOCX 124 KB)
12936_2012_2220_MOESM1_ESM.docx
Authors’ original file for figure 1
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Authors’ original file for figure 2
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Authors’ original file for figure 3
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Authors’ original file for figure 4
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