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Open Access Highly Accessed Methodology

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

Eric Marois1, Christina Scali2, Julien Soichot1, Christine Kappler1, Elena A Levashina13* and Flaminia Catteruccia45*

Author Affiliations

1 Institut de Biologie Moléculaire et Cellulaire, INSERM U963, CNRS UPR9022, 15 rue René Descartes, 67084, Strasbourg, France

2 Division of Cell and Molecular Biology, Imperial College London, Imperial College Road, London, SW7 2AZ, UK

3 Department of Vector Biology, Max-Planck Institute for Infection Biology, Chariteplatz 1, 10117, Berlin, Germany

4 Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università degli Studi di Perugia, Terni, 05100, Italy

5 Department of Immunology and Infectious Diseases, Harvard School of Public Health, 665 Huntington Avenue, Boston, MA, 02115, USA

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Malaria Journal 2012, 11:302  doi:10.1186/1475-2875-11-302

Published: 28 August 2012

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.

Keywords:
Malaria; Mosquito; Transgenesis; Fluorescence-assisted sorting; Sexing; Anopheles gambiae; piggyBac