Consequences of resistance evolution in a Cas9-based sex conversion-suppression gene drive for insect pest management
Consequences of resistance evolution in a Cas9-based sex conversion-suppression gene drive for insect pest management
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DOI：
10.1073/pnas.1713825115
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发表时间：
2018-06-12
影响因子：
11.1
通讯作者：
Wimmer, Ernst A.
中科院分区：
文献类型：
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作者：
KaramiNejadRanjbar, Mohammad;Eckermann, Kolja N.;Wimmer, Ernst A.
长期以来，人们一直在讨论使用基于位点特异性归巢的基因驱动来控制害虫，但只有随着最近CRISPR/Cas9技术的建立，这种系统的简单设计才成为可能。在这方面，关于性别决定的新发现为害虫管理提供了新的目标。在这里，我们提出了一个抑制基因驱动的模型，旨在导致农业害虫的全雄性种群崩溃。为了在实验中评估这种基于性别转换的抑制基因驱动的分子细节，我们在果蝇身上实施了这种策略，作为一种安全的模式生物。我们生成了一个基于cas9的靶向变压器基因的归巢基因驱动元件，并显示了其从雌性到雄性的高效率。然而，非同源末端连接增加了目标位点的诱变率，导致抗驱动等位基因的出现，从而抑制了基因驱动。这证实了先前的研究，即简单的归巢CRISPR/ Cas9基因驱动设计将是无效的。然而，通过使用我们在黑腹果蝇中获得的参数进行种群动态模拟，并通过调整农业害虫Ceratitis capitata模型，我们能够确定适当的修改，可以使用我们提出的基于性别转换的抑制基因驱动策略成功地应用于野生地中海果蝇种群的管理。
The use of a site-specific homing-based gene drive for insect pest control has long been discussed, but the easy design of such systems has become possible only with the recent establishment of CRISPR/Cas9 technology. In this respect, novel targets for insect pest management are provided by new discoveries regarding sex determination. Here, we present a model for a suppression gene drive designed to cause an all-male population collapse in an agricultural pest insect. To evaluate the molecular details of such a sex conversion-based suppression gene drive experimentally, we implemented this strategy in Drosophila melanogaster to serve as a safe model organism. We generated a Cas9-based homing gene-drive element targeting the transformer gene and showed its high efficiency for sex conversion from females to males. However, non-homologous end joining increased the rate of mutagenesis at the target site, which resulted in the emergence of drive-resistant alleles and therefore curbed the gene drive. This confirms previous studies that simple homing CRISPR/ Cas9 gene-drive designs will be ineffective. Nevertheless, by performing population dynamics simulations using the parameters we obtained in D. melanogaster and by adjusting the model for the agricultural pest Ceratitis capitata, we were able to identify adequate modifications that could be successfully applied for the management of wild Mediterranean fruit fly populations using our proposed sex conversion-based suppression gene-drive strategy.