When using RNA interference （RNAi） to study gene functions in Lepidoptera insects, we discovered that some genes could not be suppressed; instead, their expression levels could be up-regulated by double-stranded RNA...When using RNA interference （RNAi） to study gene functions in Lepidoptera insects, we discovered that some genes could not be suppressed; instead, their expression levels could be up-regulated by double-stranded RNA （dsRNA）. To predict which genes could be easily silenced, we treated the Asian corn borer （Ostriniafilrnacalis） with dsGFP （green fluorescent protein） and dsMLP （muscle lim protein）. A transcriptome sequence analysis was conducted using the cDNAs 6 h after treatment with dsRNA. The results indicated that 160 genes were up-regulated and 44 genes were down-regulated by the two dsRNAs. Then, 50 co-up-regulated, 25 co-down-regulated and 43 unaffected genes were selected to determine their RNAi responses. All the 25 down-regulated genes were knocked down by their corresponding dsRNA. However, several of the up-regulated and unaffected genes were up-regulated when treated with their corresponding dsRNAs instead of being knocked down. The genes up-regulated by the dsGFP treatment may be involved in insect immune responses or the RNAi pathway. When the immune-related genes were excluded, only seven genes were induced by dsGFP, including ago-2 and dicer-2. These results not only provide a reference for efficient RNAi target predications, but also provide some potential RNAi pathway-related genes for further study.展开更多
基金This work was supported by the National Basic Research Program of China （2015CB755703）, the National Transgenic Great Subject from the Ministry of Agriculture of China （2016ZX08009-003-001）, the Key Deployment Project from the Chinese Academy of Sciences （KSZD-EW-Z-021-2-1） and the National Natural Science Foundation of China （31672354 and 31402012）. The funders had no role in study design, data collection or analysis, decision to publish, or preparation of the manuscript.
文摘When using RNA interference （RNAi） to study gene functions in Lepidoptera insects, we discovered that some genes could not be suppressed; instead, their expression levels could be up-regulated by double-stranded RNA （dsRNA）. To predict which genes could be easily silenced, we treated the Asian corn borer （Ostriniafilrnacalis） with dsGFP （green fluorescent protein） and dsMLP （muscle lim protein）. A transcriptome sequence analysis was conducted using the cDNAs 6 h after treatment with dsRNA. The results indicated that 160 genes were up-regulated and 44 genes were down-regulated by the two dsRNAs. Then, 50 co-up-regulated, 25 co-down-regulated and 43 unaffected genes were selected to determine their RNAi responses. All the 25 down-regulated genes were knocked down by their corresponding dsRNA. However, several of the up-regulated and unaffected genes were up-regulated when treated with their corresponding dsRNAs instead of being knocked down. The genes up-regulated by the dsGFP treatment may be involved in insect immune responses or the RNAi pathway. When the immune-related genes were excluded, only seven genes were induced by dsGFP, including ago-2 and dicer-2. These results not only provide a reference for efficient RNAi target predications, but also provide some potential RNAi pathway-related genes for further study.
