Controlling the principal African malaria vector, the mosquito Anopheles gambiae , is considered essential to curtail malaria transmission. However, existing vector control technologies rely on insecticides, which are becoming increasingly ineffective. Sterile insect technique (SIT) is a powerful suppression approach that has successfully eradicated a number of insect pests, yet the A. gambiae toolkit lacks the requisite technologies for its implementation. SIT relies on iterative mass releases of nonbiting, nondriving, sterile males which seek out and mate with monandrous wild females. Once mated, females are permanently sterilized due to mating-induced refractoriness, which results in population suppression of the subsequent generation. However, sterilization by traditional methods renders males unfit, making the creation of precise genetic sterilization methods imperative. Here, we introduce a vector control technology termed precision-guided sterile insect technique (pgSIT), in A. gambiae for inducible, programmed male sterilization and female elimination for wide-scale use in SIT campaigns. Using a binary CRISPR strategy, we cross separate engineered Cas9 and gRNA strains to disrupt male-fertility and female-essential genes, yielding >99.5% male sterility and >99.9% female lethality in hybrid progeny. We demonstrate that these genetically sterilized males have good longevity, are able to induce sustained population suppression in cage trials, and are predicted to eliminate wild A. gambiae populations using mathematical models, making them ideal candidates for release. This work provides a valuable addition to the malaria genetic biocontrol toolkit, enabling scalable SIT-like confinable, species-specific, and safe suppression in the species.

中文翻译：

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通过精确引导的不育雄性消除疟疾媒介


控制非洲主要疟疾病媒冈比亚按蚊被认为对于遏制疟疾传播至关重要。然而，现有的病媒控制技术依赖于杀虫剂，而杀虫剂的效果越来越差。昆虫不育技术（SIT）是一种强大的抑制方法，已成功根除多种害虫，但冈比亚疟原虫工具包缺乏实施所需的技术。 SIT依赖于反复大规模释放不咬人、不驾驶、不育的雄性，这些雄性寻找单调的野生雌性并与之交配。一旦交配，雌性就会由于交配引起的难治性而永久绝育，从而导致下一代种群受到抑制。然而，传统方法的绝育会使男性变得不健康，因此创建精确的基因绝育方法势在必行。在这里，我们介绍了一种被称为精确引导昆虫不育技术 (pgSIT) 的病媒控制技术，在冈比亚 A. gambiae 中进行诱导性、程序化的雄性绝育和雌性消除，以在 SIT 活动中广泛使用。使用二元 CRISPR 策略，我们将单独的工程 Cas9 和 gRNA 菌株杂交，以破坏雄性生育力和雌性必需基因，在杂交后代中产生 >99.5% 的雄性不育性和 >99.9% 的雌性致死率。我们证明，这些基因绝育的雄性具有良好的寿命，能够在笼养试验中诱导持续的种群抑制，并且使用数学模型预测可以消除野生冈比亚种群，使它们成为释放的理想候选者。这项工作为疟疾遗传生物防治工具包提供了宝贵的补充，能够在物种中实现可扩展的类似 SIT 的有限、物种特异性和安全抑制。