Kou Zongqing, Luo Xingyu, Jiang Yuguo, Chen Bihui, Song Yu, Wang Yaohui, Xu Jun, Tomberlin Jeffery K, Huang Yongping
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, China.
University of Chinese Academy of Sciences, Beijing, China.
Insect Sci. 2023 Aug;30(4):888-900. doi: 10.1111/1744-7917.13147. Epub 2023 Jan 9.
The black soldier fly (BSF), Hermetia illucens, is a promising insect for mitigating solid waste problems as its larvae are able to bioconvert organic waste into valuable biomass. We recently reported a high-quality genome assembly of the BSF; analysis of this genome sequence will further the understanding of insect biology and identify genes that can be manipulated to improve efficiency of bioconversion. To enable genetic manipulation of the BSF, we have established the first transgenic methods for this economically important insect. We cloned and identified the ubiquitous actin5C promoter (Hiactin5C-p3k) and 3 endogenous U6 promoters (HiU6:1, HiU6:2, and HiU6:3). The Hiactin5C promoter was used to drive expression of a hyperactive variant of the piggyBac transposase, which exhibited up to 6-fold improvement in transformation rate when compared to the wild-type transposase. Furthermore, we evaluated the 3 HiU6 promoters using this transgenic system. HiU6:1 and HiU6:2 promoters provided the highest knockdown efficiency with RNAi and are thus promising candidates for future Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) development. Overall, our findings provide valuable genetic engineering toolkits for basic research and genetic manipulation of the BSF.
黑水虻（Hermetia illucens）是一种很有前景的昆虫，可用于缓解固体废物问题，因为其幼虫能够将有机废物生物转化为有价值的生物质。我们最近报道了黑水虻的高质量基因组组装；对该基因组序列的分析将进一步加深对昆虫生物学的理解，并鉴定出可进行操纵以提高生物转化效率的基因。为了实现对黑水虻的基因操纵，我们为这种具有重要经济意义的昆虫建立了首批转基因方法。我们克隆并鉴定了普遍存在的肌动蛋白5C启动子（Hiactin5C-p3k）和3个内源性U6启动子（HiU6:1、HiU6:2和HiU6:3）。Hiactin5C启动子用于驱动猪尾巴转座酶的高活性变体的表达，与野生型转座酶相比，其转化率提高了6倍。此外，我们使用该转基因系统评估了3个HiU6启动子。HiU6:1和HiU6:2启动子在RNA干扰中提供了最高的敲低效率，因此是未来成簇规律间隔短回文重复序列（CRISPR）开发的有希望的候选者。总体而言，我们的研究结果为黑水虻的基础研究和基因操纵提供了有价值的基因工程工具包。
