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Molecular characterization and RNA interference responses of the lethal giant larvae gene in Diabrotica virgifera virgifera adults
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2021-04-19 , DOI: 10.1002/arch.21787 Anastasia M. W. Cooper 1 , Huifang Song 1, 2 , Xuekai Shi 1, 2 , Zhitao Yu 1 , Young Ho Kim 1, 3 , Kristopher Silver 1 , Jianzhen Zhang 1, 2 , Kun Yan Zhu 1
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2021-04-19 , DOI: 10.1002/arch.21787 Anastasia M. W. Cooper 1 , Huifang Song 1, 2 , Xuekai Shi 1, 2 , Zhitao Yu 1 , Young Ho Kim 1, 3 , Kristopher Silver 1 , Jianzhen Zhang 1, 2 , Kun Yan Zhu 1
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High specificity for silencing target genes and single-copy target genes that yield clear phenotypes are two important factors for the success of RNA interference (RNAi). The lethal giant larvae (Lgl) gene appears to be an ideal gene for RNAi because RNAi can effectively suppress its expression and results in molting defects and mortality in Tribolium castaneum. To investigate the suitability of this gene for RNAi in other insects, we identified and characterized DvLgl from the western corn rootworm, Diabrotica virgifera virgifera, a species exhibiting high RNAi efficiency. DvLgl was expressed in all developmental stages and tissues investigated. The deduced DvLgl protein showed high amino-acid sequence identities and similar domain architecture to Lgls from other insect species. Despite many similarities among insect Lgls, RNAi-mediated suppression of DvLgl failed to produce a phenotype in D. v. virgifera adults. The difference in developing phenotypes could be attributed greatly to the level of gene suppression and the insect developmental stages for RNAi. These results highlight the variability in RNAi response among insects and showcase the importance of screening multiple target genes when conducting RNAi studies. Our findings are expected to help the design of future RNAi studies and future investigations of Lgl in insects.
中文翻译：
致命假蝇幼虫基因的分子表征和RNA干扰响应
沉默靶基因和产生清晰表型的单拷贝靶基因的高度特异性是成功实现RNA干扰（RNAi）的两个重要因素。致死巨型幼虫（LGL）基因似乎是RNAi的一个理想的基因，因为RNAi可有效地抑制在蜕皮缺陷和死亡率其表达并导致赤拟谷盗。为了研究该基因在其他昆虫中对RNAi的适应性，我们从西部玉米根虫Diabrotica virgifera virgifera鉴定并鉴定了DvLgl，该物种显示出较高的RNAi效率。DvLgl在研究的所有发育阶段和组织中均表达。推导的DvLgl蛋白显示出很高的氨基酸序列同一性，并且结构域结构与其他昆虫的Lgl类似。尽管昆虫Lgls之间有许多相似之处，但RNAi介导的DvLgl抑制在D. v。virgifera成虫中未产生表型。发育表型的差异可归因于RNAi的基因抑制水平和昆虫发育阶段。这些结果凸显了昆虫之间RNAi反应的变异性，并展示了进行RNAi研究时筛选多个靶基因的重要性。预期我们的发现将有助于设计未来的RNAi研究和昆虫中Lgl的未来研究。
更新日期：2021-05-26
中文翻译：
致命假蝇幼虫基因的分子表征和RNA干扰响应
沉默靶基因和产生清晰表型的单拷贝靶基因的高度特异性是成功实现RNA干扰（RNAi）的两个重要因素。致死巨型幼虫（LGL）基因似乎是RNAi的一个理想的基因，因为RNAi可有效地抑制在蜕皮缺陷和死亡率其表达并导致赤拟谷盗。为了研究该基因在其他昆虫中对RNAi的适应性，我们从西部玉米根虫Diabrotica virgifera virgifera鉴定并鉴定了DvLgl，该物种显示出较高的RNAi效率。DvLgl在研究的所有发育阶段和组织中均表达。推导的DvLgl蛋白显示出很高的氨基酸序列同一性，并且结构域结构与其他昆虫的Lgl类似。尽管昆虫Lgls之间有许多相似之处，但RNAi介导的DvLgl抑制在D. v。virgifera成虫中未产生表型。发育表型的差异可归因于RNAi的基因抑制水平和昆虫发育阶段。这些结果凸显了昆虫之间RNAi反应的变异性，并展示了进行RNAi研究时筛选多个靶基因的重要性。预期我们的发现将有助于设计未来的RNAi研究和昆虫中Lgl的未来研究。
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