Establishing a dominant early larval sex-selection strain in the Asian malaria vector Anopheles stephensi  

作　　者：Shih‑Che Weng Fangying Chen Ming Li Sammy Lee Connor Gerry Dylan Can Turksoy Omar S.Akbari 

机构地区：[1]School of Biological Sciences,Department of Cell and Developmental Biol‑ogy,University of California,San Diego,La Jolla,CA 92093,USA

出　　处：《Infectious Diseases of Poverty》2024年第6期80-89,共10页贫困所致传染病(英文)

基　　金：supported by funding from NIH awards(R01AI151004,RO1AI148300,RO1AI175152)awarded to O.S.A.Figures were created using Prism9 for Windows(GraphPad Software,San Diego,USA)and www.BioRe nder.com.

摘　　要：Background Genetic biocontrol interventions targeting mosquito-borne diseases require the release of male mosquitoes exclusively,as only females consume blood and transmit pathogens.Releasing only males eliminates the risk of increasing mosquito bites and spreading pathogens while enabling effective population control.The aim of this study is to develop robust sex-sorting methods for early larval stages in mosquitoes,enabling scalable male-only releases for genetic biocontrol interventions.Methods To address the challenge of sex-sorting in the Asian malaria vector Anopheles stephensi,we engineer Sexing Element Produced by Alternative RNA-splicing of a Transgenic Observable Reporter(SEPARATOR).This dominant fluorescent-based method,previously proven effective in Aedes aegypti,exploits sex-specifiic alternative splicing of a reporter to ensure exclusive male-specific expression early in development.The sex-specific alternative RNA splicing of the doublesex gene was selected as a target for engineering SEPARATOR due to its evolutionary conservation in insects.To expand SEPARATOR's applicability for genetic sexing,we assessed the cross-species sex-specific RNA splicing activity of the An.gambiae doublesex(AngDsx)splicing module in An.stephensi.Male-specific enhanced green fluorescent protein(EGFP)expression was verified throughout the mosquito life cycle using a fluorescent stereomicroscope.Results Our results confirm that SEPARATOR regulates male-specific EGFP expression in An.stephensi and enables reliable positive male selection from the first instar larval stages.Molecular analysis demonstrates that male-specific EGFP expression is dependent on doublesex sex-specific splicing events.Additionally,the splicing module from An.gambiae operates effectively in An.stephensi,demonstrating evolutionary conservation in sex-specific splicing events between these species.Conclusions SEPARATOR's independence from sex-chromosome linkage provides resistance to breakage that could be mediated by meiotic recombination and chromos

关 键 词：Genetic sex sorting Malaria DOUBLESEX Sex-specific alternative splicing Anopheles stephensi Genetic biocontrol 

分 类 号：G63[文化科学—教育学]