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鉴定、功能表征和系统进化分析沙漠蝗肠道中的双链 RNA 降解酶。

Identification, functional characterization and phylogenetic analysis of double stranded RNA degrading enzymes present in the gut of the desert locust, Schistocerca gregaria.

机构信息

Molecular Developmental Physiology and Signal Transduction, KU Leuven, Naamsestraat 59, P.O. Box 02465, B-3000 Leuven, Belgium.

Molecular Developmental Physiology and Signal Transduction, KU Leuven, Naamsestraat 59, P.O. Box 02465, B-3000 Leuven, Belgium.

出版信息

Insect Biochem Mol Biol. 2014 Mar;46:1-8. doi: 10.1016/j.ibmb.2013.12.008. Epub 2014 Jan 10.

Abstract

RNA interference (RNAi) has become a widely used reverse genetics tool in eukaryotes and holds great potential to contribute to the development of novel strategies for insect pest control. While previous studies clearly demonstrated that injection of dsRNA into the body cavity of the desert locust, Schistocerca gregaria, is highly effective to induce gene silencing effects, we observed that the RNAi response is much less sensitive to orally delivered dsRNA. In line with this, we report on the presence of a potent dsRNA degrading activity in the midgut juice. Four different dsRNase sequences that belong to the DNA/RNA Non-specific Nuclease superfamily were retrieved from a transcriptome database of the desert locust. Surprisingly, we have found that, in the publicly available eukaryote nucleotide sequence databases, the presence of this group of enzymes is restricted to insects and crustaceans. Nonetheless, phylogenetic analyses predict a common origin of these enzymes with the Endonuclease G (EndoG) Non-specific Nucleases that display a widespread taxonomic distribution. Moreover, in contrast to the Sg-endoG transcript, the four Sg-dsRNase transcripts appear to be specifically expressed in the gut. Finally, by means of RNAi, we provide evidence for an important contribution of dsRNase2 to the dsRNA degrading activity that is present in the gut lumen of S. gregaria.

摘要

RNA 干扰 (RNAi) 已成为真核生物中广泛使用的反向遗传学工具,对于开发新型昆虫防治策略具有很大的潜力。虽然先前的研究清楚地表明,将 dsRNA 注射到沙漠蝗的体腔中高度有效地诱导基因沉默效应,但我们观察到 RNAi 反应对口服给予的 dsRNA 的敏感性要低得多。与此一致,我们报告了中肠汁液中存在一种有效的 dsRNA 降解活性。从沙漠蝗的转录组数据库中检索到属于 DNA/RNA 非特异性核酸酶超家族的四种不同的 dsRNase 序列。令人惊讶的是,我们发现,在公开的真核生物核苷酸序列数据库中,这组酶的存在仅限于昆虫和甲壳类动物。尽管如此,系统发育分析预测这些酶与具有广泛分类分布的内切核酸酶 G(EndoG)非特异性核酸酶具有共同的起源。此外,与 Sg-endoG 转录物相比,四种 Sg-dsRNase 转录物似乎专门在肠道中表达。最后,通过 RNAi,我们提供了证据表明 dsRNase2 对存在于 S. gregaria 肠腔中的 dsRNA 降解活性有重要贡献。

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