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您的检索式：作者名="Kristopher Silver"
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| 1 | Identification of Rab family genes and functional analyses of LmRab5 and LmRab11A in the development and RNA interference of Locusta migratoria显示文摘Rab proteins constitute the largest family of small GTPases,which play pivotal roles in intracellular membrane trafficking in all eukaryotes.A number of Rab genes have been identified in eukaryotes;however,very little information about these genes has been reported in insects.In the current study,for the first time we identified and characterized 27 Rab family genes from Locusta migratoria.Phylogenetic analysis and comparison of domain architecture indicated that Rab family genes are highly conserved among insect species.Tissue-dependent expression profiles indicated that expression of Rab genes was highest in the ovary,except for LmRab3,which was most highly expressed in hemolymph.The biological function of each Rab gene was investigated using RNA interference(RNAi).Double-stranded RNA targeting each Rab gene was injected into the hemocoel of nymphs and revealed that suppression of two Rab genes(LmRab5 and LmRab11A)caused 100%mortality.In addition,nymphs injected with dsLmRab5 exhibited severe phenotypic defects in the gastric caeca and midgut,while dsLmRab11A arrested the molting process.We then applied the RNAi of RNAi technique to test if silencing either of these two genes would affect the suppression of the lethal giant larvae(LmLgl)reporter gene and found that suppression of LmRab5 diminished the RNAi efficiency of LmLgl,whereas suppression of LmRab11A enhanced RNAi efficiency of LmLgl.These results indicate that Rab genes contribute differently to RNAi efficiency in different tissues.Our study provides a foundation for further functional investigations of Rab genes and their contributions to RNAi efficiency in L.migratoria. | Mureed Abbas Yun-He Fan Xue-Kai Shi Lu Gao Yan-Li Wang Tao Li Anastasia M.W.Cooper Kristopher Silver Kun Yan Zhu Jian-Zhen Zhang | 2022 | Insect Science2022，29，2： | 0 |
| 2 | Lethal giant larvae gene is required for normal nymphal development and midgut morphogenesis in Locusta migratoria显示文摘Disruption of morphogenesis,an essential process in organismal development,can lead to disruption of biological processes,reduction in fitness,or even death of an organism.The roles of lethal giant larvae(Lgl)protein in maintaining tissue organization have been studied extensively in mammals,but little is known about this gene's roles in promoting correct tissue morphogenesis in insects.In this study,we identified an Lgl ortholog in Locusta migratoria.RT-qPCR results revealed that LmLgl was constitutively expressed during third,fourth,and fifth instar nymphs.Furthermore,LmLgl showed highest expression in the ovary followed by wing pads,midgut,hindgut,Malpighian tubules,and foregut of the third-instar nymphs.To examine the role of LmLgl in L.migratoria development,RNA interference was performed during nymphal stages.Silencing of LmLgl increased body size but decreased bodyweight by 9.0%.Histological sections of the midgut revealed abnormal large masses of disordered epithelial cells in dsLmLgl-injected nymphs.In addition,downregulation of LmLgl transcript levels significantly altered the morphological structure in midgut,resulting in the formation of tumor-like structures.Our results indicated that LmLgl may act as a tumor-suppressor gene,which plays an essential role in maintaining a normal morphological structure in the midgut of L.migratoria.Our results also suggest that LmLgl may be explored as a potential target for developing dsRNA-based biological pesticides for managing insect pests. | Xuekai Shi Xiaojian Liu Kristopher Silver Kun Yan Zhu Jianzhen Zhang | 2022 | Insect Science2022，29，4： | 0 |
| 3 | A dsRNA-degrading nuclease(dsRNase2)limits RNAi efficiency in the Asian corn borer(Ostrinia furnacalis)显示文摘The efficiency of RNA interference(RNAi)varies substantially among different insect species.Rapid degradation of double-stranded RNA(dsRNA)by dsRNA-degrading nucleases(dsRNases)has been implicated to cause low RNAi efficiency in several insect species.In this study,we identified four dsRNase genes(OfdsRNaseL Ofd-sRNase2,OfdsRNase3 and OfdsRNase4)from the Asian corn borer(Ostrinia furnacalis)transcriptome database.Bioinformatic analyses showed that each deduced protein sequence contained endonuclease NS domains and signal peptides.Gene expression analysis revealed that OfdsRNase2 was exclusively expressed in the midgut of larvae.RNAi efficiency was investigated in 2-d-old fifth-instar larvae(high expression of dsRNase2)and 2-d-old pupae(low expression of dsRNase2)by feeding or injecting dsRNA targeting a marker gene that encodes the lethal giant larvae protein(OfLgl).Our results showed that OfLgl only partially silenced the expression of OfLgl in pupae,but not in larvae,suggesting that OfdsRNase2 could contribute to lower RNAi efficiency in larval stages.This hypothesis was supported by our RNAi-of-RNAi experiment using a tissue culture technique where the silencing efficiency against the reporter gene,OfHexl,was significantly improved after knockdown of OfdsRNase2.When double luciferase assays were performed to evaluate the role of the four dsRNases in vitro,only OfdsRNase2 expressed in S2 cells significantly affected RNAi efficiency by degrading dsRNA.Taken together,our results suggested that the degradation of dsRNA by OfdsRNase2 in the midgut contributed to low RNAi efficiency in O.furnacalis larvae. | Yun-He Fan Hui-Fang Song Mureed Abbas Yan-Li Wang Tao Li En-Bo Ma Anastasia M.W.Cooper Kristopher Silver Kun Yan Zhu Jian-Zhen Zhang | 2021 | Insect Science2021，28，6： | 0 |
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