Kou Zongqing, Wang Shaozhen, Luo Xingyu, Xu Jun, Tomberlin Jeffery K, Huang Yongping
School of Environmental Science and Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
Key Laboratory of Insect Developmental and Evolutionary Biology, Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, 200032, China.
Commun Biol. 2024 Dec 19;7(1):1652. doi: 10.1038/s42003-024-07254-7.
Larvae of the black soldier fly Hermetia illucens have potential as a natural waste recycler and subsequent use as protein-rich feed for livestock. A common question about the insect-farming processes is, what about the concerns of mass escape of insects from large populations? Here, we present a binary transgenic CRISPR/Cas9 system to generate wingless strain with the potential to address this issue. We identified gonad-specific promoters in vivo and evaluated use of the two strongest promoters, nanos and exuperantia, to drive Cas9 expression. We found that crossing the Hiexu-Cas9 with transgenic sgRNA-expressing insects resulted in higher knockout efficiency of the marker gene white. The Hiexu-Cas9 strain exhibited a maternal deposition of Cas9 that caused more effective knockout in the progeny of female Cas9-expressing individuals. Using this system, we generated wingless mutants lacking mating ability, which can be maintained in colony through a genetic cross of two single strain. These insects are less likely to escape and would be unable to successfully mate if they did escape. Taken together, this study validates effective genetic tools that can be used for gene function studies and industrial applications in black soldier fly and provides an approach to alleviate the concern about massive rearing.
黑水虻（Hermetia illucens）幼虫有潜力成为天然的废物回收者，并随后用作富含蛋白质的家畜饲料。关于昆虫养殖过程的一个常见问题是，大量昆虫从大规模养殖群体中逃逸的担忧该如何解决？在此，我们展示了一种二元转基因CRISPR/Cas9系统，以产生无翅品系，有潜力解决这一问题。我们在体内鉴定了性腺特异性启动子，并评估了使用两个最强的启动子——nanos和exuperantia来驱动Cas9表达。我们发现，将Hiexu-Cas9与表达转基因sgRNA的昆虫杂交，导致标记基因白色的敲除效率更高。Hiexu-Cas9品系表现出Cas9的母系沉积，这在表达Cas9的雌性个体的后代中导致更有效的敲除。使用这个系统，我们产生了缺乏交配能力的无翅突变体，它们可以通过两个单菌株的遗传杂交在群体中维持。这些昆虫逃逸的可能性较小，并且如果它们确实逃逸，也无法成功交配。综上所述，本研究验证了可用于黑水虻基因功能研究和工业应用的有效遗传工具，并提供了一种减轻对大规模饲养担忧的方法。
