Southworth Joshua, Gonzalez Estela, Nevard Katherine, Larrosa-Godall Mireia, Alphey Luke, Anderson Michelle A E
Department of Biosciences, Durham University, Durham, UK.
Arthropod Genetics, The Pirbright Institute, Pirbright, UK.
Insect Mol Biol. 2025 Feb;34(1):104-110. doi: 10.1111/imb.12953. Epub 2024 Aug 11.
Anopheles stephensi Liston, 1901 (Diptera: culicidae) is a competent vector of Plasmodium falciparum (Haemosporida: plasmodiidae) malaria, and its expansion in the African continent is of concern due to its viability in urban settings and resistance to insecticides. To enhance its genetic tractability, we determined the utility of a ~2 kb An. stephensi lipophorin (lp) promoter fragment in driving transgene expression. Lipophorin genes are involved in lipid transport in insects, and an orthologous promoter in An. gambiae (AGAP001826) was previously demonstrated to successfully express a transgene. In the present study, we qualitatively characterised the expression of a ZsYellow fluorescent marker protein, expressed by An. stephensi lp promoter fragment. Our study indicated that the lp promoter fragment was effective, generating a distinct expression pattern in comparison to the commonly utilised 3xP3 promoter. The lp:ZsYellow fluorescence was largely visible in early instar larvae and appeared more intense in later instar larvae, pupae and adults, becoming especially conspicuous in adult females after a blood meal. Different isolines showed some variation in expression pattern and intensity. Aside from general transgene expression, as the lp promoter produces a suitable fluorescent protein marker expression pattern, it may facilitate genotypic screening and aid the development of more complex genetic biocontrol systems, such as multi-component gene drives. This study represents an expansion of the An. stephensi genetic toolbox, an important endeavour to increase the speed of An. stephensi research and reach public health milestones in combating malaria.
斯氏按蚊（Anopheles stephensi Liston，1901年）（双翅目：蚊科）是恶性疟原虫（血孢子虫目：疟原虫科）疟疾的有效传播媒介，由于其在城市环境中的生存能力以及对杀虫剂的抗性，它在非洲大陆的扩散令人担忧。为了提高其遗传可操作性，我们确定了一个约2 kb的斯氏按蚊脂运载蛋白（lp）启动子片段在驱动转基因表达方面的效用。脂运载蛋白基因参与昆虫体内的脂质运输，此前已证明冈比亚按蚊（AGAP001826）中的一个直系同源启动子能够成功表达转基因。在本研究中，我们对由斯氏按蚊lp启动子片段表达的ZsYellow荧光标记蛋白的表达进行了定性表征。我们的研究表明，lp启动子片段是有效的，与常用的3xP3启动子相比，产生了独特的表达模式。lp:ZsYellow荧光在早期幼虫中基本可见，在后期幼虫、蛹和成虫中显得更强，在成年雌性按蚊吸血后尤其明显。不同的同基因系在表达模式和强度上表现出一些差异。除了一般的转基因表达外，由于lp启动子产生了合适的荧光蛋白标记表达模式，它可能有助于基因型筛选，并有助于开发更复杂的遗传生物防治系统，如多组分基因驱动。本研究代表了斯氏按蚊遗传工具箱的扩展，这是加快斯氏按蚊研究速度并在抗击疟疾方面实现公共卫生里程碑的一项重要努力。
