在亚洲疟疾病媒疟原虫中建立优势早期幼虫性别选择株系。

IF 8.1 1区 医学
Shih-Che Weng, Fangying Chen, Ming Li, Sammy Lee, Connor Gerry, Dylan Can Turksoy, Omar S Akbari
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引用次数: 0

摘要

背景:针对蚊媒疾病的基因生物控制干预措施要求只释放雄蚊,因为只有雌蚊才吸血和传播病原体。只释放雄蚊可以消除增加蚊虫叮咬和传播病原体的风险,同时实现有效的种群控制。这项研究的目的是为蚊子的早期幼虫阶段开发可靠的性别分选方法,使基因生物控制干预中的只释放雄性成为可能:方法:为了解决亚洲疟疾病媒申氏疟蚊性别分类的难题,我们设计了通过替代 RNA 剪接转基因可观察报告(SEPARATOR)产生的性别元素。这种基于显性荧光的方法以前曾在埃及伊蚊中被证明是有效的,它利用报告基因的性别特异性替代剪接来确保在发育早期只表达雄性特异性。由于在昆虫的进化过程中,双性基因的性别特异性替代 RNA 剪接一直被视为 SEPARATOR 工程的目标。为了扩大 SEPARATOR 在遗传性别鉴定中的适用性,我们评估了冈比亚蚁双性基因(AngDsx)剪接模块在史蒂芬蚁(An. stephensi)中的跨物种性别特异性 RNA 剪接活性。使用荧光体视显微镜验证了蚊子整个生命周期中雄性特异性增强绿色荧光蛋白(EGFP)的表达:结果:我们的研究结果证实,SEPARATOR调控了史蒂芬蚊的雄性特异性EGFP表达,并从第一龄幼虫阶段就实现了可靠的阳性雄性选择。分子分析表明,雄性特异性 EGFP 表达依赖于双倍性性别特异性剪接事件。此外,冈比亚蚁的剪接模块在史蒂芬蚁中也能有效运作,这表明这些物种之间的性别特异性剪接事件在进化上是一致的:SEPARATOR独立于性染色体连接,可抵御由减数分裂重组和染色体重排引起的断裂,因此非常适合大规模雄性释放。SEPARATOR 能够从第一龄幼虫阶段就开始进行精确的雄性选择,是一项重大进步,将有助于对疟蚊进行遗传生物控制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Establishing a dominant early larval sex-selection strain in the Asian malaria vector Anopheles stephensi.

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-specific 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 chromosomal rearrangements, making it highly suitable for mass male releases. By enabling precise male selection from the first instar larval stages, SEPARATOR represents a significant advancement that will aid in the genetic biocontrol for Anopheles mosquitoes.

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来源期刊
Infectious Diseases of Poverty
Infectious Diseases of Poverty INFECTIOUS DISEASES-
自引率
1.20%
发文量
368
期刊介绍: Infectious Diseases of Poverty is an open access, peer-reviewed journal that focuses on addressing essential public health questions related to infectious diseases of poverty. The journal covers a wide range of topics including the biology of pathogens and vectors, diagnosis and detection, treatment and case management, epidemiology and modeling, zoonotic hosts and animal reservoirs, control strategies and implementation, new technologies and application. It also considers the transdisciplinary or multisectoral effects on health systems, ecohealth, environmental management, and innovative technology. The journal aims to identify and assess research and information gaps that hinder progress towards new interventions for public health problems in the developing world. Additionally, it provides a platform for discussing these issues to advance research and evidence building for improved public health interventions in poor settings.
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