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

Malaria Journal

Methodology

Flaminia Catteruccia¹  Christina Scali²  Christine Kappler³  Julien Soichot³  Eric Marois³  Elena A Levashina⁴ 

[1] Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università degli Studi di Perugia, 05100, Terni, Italy;Department of Immunology and Infectious Diseases, Harvard School of Public Health, 665 Huntington Avenue, 02115, Boston, MA, USA;Division of Cell and Molecular Biology, Imperial College London, Imperial College Road, SW7 2AZ, London, UK;Institut de Biologie Moléculaire et Cellulaire, INSERM U963, CNRS UPR9022, 15 rue René Descartes, 67084, Strasbourg, France;Institut de Biologie Moléculaire et Cellulaire, INSERM U963, CNRS UPR9022, 15 rue René Descartes, 67084, Strasbourg, France;Department of Vector Biology, Max-Planck Institute for Infection Biology, Chariteplatz 1, 10117, Berlin, Germany;

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

DOI  :  10.1186/1475-2875-11-302

 received in 2012-05-25, accepted in 2012-08-07,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundMosquito 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.MethodsA 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.ResultsFluorescence-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.ConclusionsThe 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.

【 授权许可】

Unknown   
© Marois et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.