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

• Published in: Malaria journal Vol. 11; no. 1; p. 302
• Main Authors: Marois, Eric, Scali, Christina, Soichot, Julien, Kappler, Christine, Levashina, Elena A, Catteruccia, Flaminia
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 28.08.2012; BioMed Central; BMC
• Subjects: Agreements; Animals; Animals, Genetically Modified - classification; Animals, Genetically Modified - genetics; Anopheles; Anopheles - classification; Anopheles - genetics; Anopheles gambiae; Biology; Communicable diseases; Competition; Control; Development and progression; Disease; Disease transmission; Entomology - methods; Female; Females; Flow Cytometry - methods; Fluorescence-assisted sorting; Gene expression; Genes, Reporter; Genetic engineering; Genetic vectors; Genetics; Genomes; Genotype; High-Throughput Screening Assays - methods; Human health and pathology; Humans; Infectious diseases; Insecticides; Larva - classification; Larva - genetics; Life Sciences; Malaria; Male; Males; Methodology; Mosquito; Mosquitoes; piggyBac; Sensitivity and Specificity; Sex; Sexing; Sperm; Transgenesis; Tropical diseases; Malaria; Anopheles gambiae; Mosquito; Sexing; Fluorescence-assisted sorting; piggyBac; Transgenesis
• ISSN: 1475-2875; 1475-2875
• DOI: 10.1186/1475-2875-11-302

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. 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. 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. 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.