When using RNA interference (RNAi) to study gene functions in Lepidopterainsects,we discovered that some genes could not be suppressed;instead,their expressionlevels could be up-regulated by double-stranded RNA (dsRNA).To predict which genescould be easily silenced,we treated the Asian co borer (Ostriniafuacalis) with dsGFP(green fluorescent protein) and dsMLP (muscle lim protein).A transcriptome sequenceanalysis was conducted using the cDNAs 6 h after treatment with dsRNA.The resultsindicated that 160 genes were up-regulated and 44 genes were down-regulated by the twodsRNAs.Then,50 co-up-regulated,25 co-down-regulated and 43 unaffected genes wereselected to determine their RNAi responses.All the 25 down-regulated genes were knockeddown by their corresponding dsRNA.However,several of the up-regulated and unaffectedgenes were up-regulated when treated with their corresponding dsRNAs instead of beingknocked down.The genes up-regulated by the dsGFP treatment may be involved in insectimmune 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 notonly provide a reference for efficient RNAi target predications,but also provide somepotential RNAi pathway-related genes for further study.