Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
Insect Mol Biol. 2021 Aug;30(4):420-426. doi: 10.1111/imb.12707. Epub 2021 May 24.
The invasive spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) has caused serious economic losses to the fruit industry. The conventional control methods have many limitations and genetic engineering technologies such as CRISPR/Cas9-mediated gene drive are promising approaches. In the CRISPR/Cas9 system, the transcriptional regulatory elements play an important role in the activities of gRNA. Thus, in order to improve the genome editing efficiency of the CRISPR/Cas9 system in D. suzukii, we cloned and tested four endogenous U6 promoters to drive mutagenesis of the white gene. Our results showed that all the four promoters could be used with variable efficiency. The promoter DsU6-3 had the highest genome editing efficiency among the four DsU6 promoters. Compared with the DsU6-3 promoter, the DmU6:3 promoter showed lower efficiency to drive mutagenesis in D. suzukii. These findings expand the range of promoters available to express gRNAs in D. suzukii, facilitating the basic and applied research on this important pest.
入侵性斑翅果蝇 Drosophila suzukii（松田果蝇）（双翅目：果蝇科）对水果产业造成了严重的经济损失。传统的控制方法有许多局限性，而 CRISPR/Cas9 介导的基因驱动等遗传工程技术则是很有前途的方法。在 CRISPR/Cas9 系统中，转录调控元件在 gRNA 的活性中起着重要作用。因此，为了提高 CRISPR/Cas9 系统在 D. suzukii 中的基因组编辑效率，我们克隆并测试了四个内源性 U6 启动子来驱动白基因的突变。结果表明，这四个启动子都可以在不同的效率下使用。四个 DsU6 启动子中，DsU6-3 启动子的基因组编辑效率最高。与 DsU6-3 启动子相比，DmU6:3 启动子在驱动 D. suzukii 突变方面的效率较低。这些发现扩展了可在 D. suzukii 中表达 gRNA 的启动子范围，为该重要害虫的基础和应用研究提供了便利。
