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Comparison of effects of dsRNA and siRNA RNA interference on insulin-like androgenic gland gene (IAG) in red swamp crayfish Procambarus clarkii

文献类型： 外文期刊

作者： Ge, Hai-Lun 1 ; Tan, Kianann 1 ; Shi, Lin-Lin 1 ; Sun, Rong 1 ; Wang, Wei-Min 1 ; Li, Yan-He 1 ;

作者机构： 1.Huazhong Agr Univ, Coll Fisheries, Key Lab Agr Anim Genet Breeding & Reprod, Minist Educ,Key Lab Freshwater Anim Breeding,Mini, Wuhan, Peoples R China

2.Chinese Acad Fishery Sci, Freshwater Fisheries Res Ctr, Key Lab Genet Breeding & Aquaculture Biol Freshwa, Minist Agr, Wuxi 214081, Jiangsu, Peoples R China

关键词： RNA interference; Procambarus clarkii; IAG; DsRNA; SiRNA

期刊名称：GENE （ 影响因子：3.688； 五年影响因子：3.329 ）

ISSN： 0378-1119

年卷期： 2020 年 752 卷

页码：

收录情况： SCI

摘要： RNA interference (RNAi), which employs double-strand RNA (dsRNA) or small interference RNA (siRNA), is a popular reverse genetic manipulation tool to study gene function. Presently, there is few reports on the implementation of RNAi on the insulin-like androgenic gland gene (MG) in red swamp crayfish Procambarus clarkii. In this study, the effective sequence of siRNA and optimal injection dose were determined, and the effects of RNAi using dsRNA, siRNA, and long-term RNAi were investigated. The results showed that the doses of 0.5 and 1 mu g/g of body weight of IAG-siRNA3 produced significantly better inhibition than 0.1 mu g/g. qPCR assays showed that both dsRNA and siRNA silenced the IAG expression in five tissues (brain, ventral nerve cord, androgenic gland, testis, and vas deferens) in adult P. clarkii, with the effectiveness decreasing over time, inhibiting the production of spermatid. dsRNA exhibited a longer interference effect than siRNA in adults. For long-term interference (P. clarkii juveniles were injected 7 times with 1 mu g/g of body weight of IAG-dsRNA), and found that the secondary sexual characteristics of juveniles were affected, while the control group developed normally. The results of this study could lay the foundation for crayfish sex reversal with MG RNAi, and provide the reference for those studies in which the technique of RNAi was used.

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