当前位置: X-MOL 学术Pestic. Biochem. Phys. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Stability of double-stranded RNA in gut contents and hemolymph of Ostrinia nubilalis larvae
Pesticide Biochemistry and Physiology ( IF 4.7 ) Pub Date : 2020-10-01 , DOI: 10.1016/j.pestbp.2020.104672
Anastasia M W Cooper 1 , Zhitao Yu 1 , Marie Biondi 1 , Huifang Song 2 , Kristopher Silver 1 , Jianzhen Zhang 2 , Kun Yan Zhu 1
Affiliation  

RNA interference (RNAi) is a revolutionary technique for silencing gene expression, but the success of this technique is dependent upon the stability of double-stranded RNA (dsRNA) molecules. In many insects, especially lepidopteran species, RNAi efficiency is limited by high instability of dsRNA in the gut and/or hemolymph, preventing the development of RNAi-based strategies for many serious pests. Previous attempts to perform RNAi on Ostrinia nubilalis (ECB, Lepidoptera: Crambidae) indicate low RNAi efficiency with both dsRNA injection and feeding. To investigate the contribution of dsRNA instability to low RNAi efficiency in ECB, a serious of ex vivo incubation experiments were performed where dsRNA integrity was assessed following incubation in larval gut continents and hemolymph using gel electrophoresis or RT-qPCR. DsRNA was less stable in the gut contents from ECB than in gut contents from Diabrotica virgifera virgifera, a coleopteran exhibiting high RNAi efficiency. Furthermore, characterization of dsRNA stability in ECB gut contents and hemolymph revealed that dsRNA was rapidly degraded under physiologically relevant conditions as a result of enzymatic activity that was neither size- nor sequence-dependent. These findings suggest that instability of dsRNA in ECB tissues is a contributing factor to the poor efficiency of RNAi in this pest. This work advances our understanding of mechanisms impacting RNAi efficiency in ECB and related lepidopteran insects for which novel pest management strategies are needed, and may facilitate the development of strategies for enhancing dsRNA stability in ECB tissues.

中文翻译:

双链RNA在Ostrinia nubilalis幼虫肠道内容物和血淋巴中的稳定性

RNA 干扰 (RNAi) 是一种革命性的基因表达沉默技术,但该技术的成功取决于双链 RNA (dsRNA) 分子的稳定性。在许多昆虫中,尤其是鳞翅目物种,RNAi 效率受到肠道和/或血淋巴中 dsRNA 的高度不稳定性的限制,从而阻碍了针对许多严重害虫的基于 RNAi 的策略的发展。以前对 Ostrinia nubilalis(ECB,鳞翅目:蟹科)进行 RNAi 的尝试表明,dsRNA 注射和喂养的 RNAi 效率较低。为了研究 d​​sRNA 不稳定性对 ECB 中低 RNAi 效率的贡献,进行了一系列离体孵育实验,其中使用凝胶电泳或 RT-qPCR 在幼虫肠道大陆和血淋巴中孵育后评估了 dsRNA 完整性。与来自 Diabrotica virgifera virgifera(一种表现出高 RNAi 效率的鞘翅目昆虫)的肠道内容物相比,来自 ECB 的肠道内容物中的 DsRNA 更不稳定。此外,对 ECB 肠道内容物和血淋巴中 dsRNA 稳定性的表征表明,由于既不依赖于大小也不依赖于序列的酶活性,dsRNA 在生理相关条件下会迅速降解。这些发现表明,ECB 组织中 dsRNA 的不稳定性是导致该害虫 RNAi 效率低下的一个因素。这项工作促进了我们对影响 ECB 和相关鳞翅目昆虫 RNAi 效率的机制的理解,这些昆虫需要新的害虫管理策略,并可能有助于制定增强 ECB 组织中 dsRNA 稳定性的策略。
更新日期:2020-10-01
down
wechat
bug