Comparison of strategies for enhancing RNA interference efficiency in Ostrinia nubilalis.
Comparison of strategies for enhancing RNA interference efficiency in Ostrinia nubilalis.
复制标题
增强玉米螟 RNA 干扰效率的策略比较。
DOI：
10.1002/ps.6114
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发表时间：
2021-03
影响因子：
4.1
通讯作者：
Zhu KY
中科院分区：
文献类型：
--
作者：
Cooper AM;Song H;Yu Z;Biondi M;Bai J;Shi X;Ren Z;Weerasekara SM;Hua DH;Silver K;Zhang J;Zhu KY
利用RNA干扰（RNAi）技术沉默昆虫特异性基因是一种新的害虫防治策略。然而，鳞翅目昆虫对RNAi是排斥的，这阻碍了新的RNAi技术应用于许多臭名昭著的害虫，包括欧洲玉米螟（Ostrinia nubilalis，ECB）。用于增强RNAi效率的策略，包括大剂量的双链RNA（dsRNA）、核酸酶抑制剂、转染试剂和纳米颗粒，已经证明可用于表现出大量dsRNA降解的其他昆虫，这是限制RNAi功效的主要机制。为了确定类似的策略是否可以增强ECB中的RNAi效率，测试了各种试剂在ECB组织中增强dsRNA稳定性的能力，然后比较了它们在整个ECB中的有效性。离体孵育实验显示，与未涂覆的dsRNA相比，Metafectene Pro、EDTA、基于壳聚糖的dsRNA纳米颗粒和Zn 2+增强了dsRNA在ECB血淋巴和肠内容物提取物中的稳定性。尽管有这些积极的结果，但本研究中使用的试剂在体内增强ECB中的RNAi效率方面是无效的。为了减少测定时间和所需的dsRNA，解剖中肠并在含有dsRNA的组织培养基中孵育，有和没有试剂。这些实验表明，RNAi效率在靶基因之间变化，并且核酸酶抑制剂仅改善了离体研究的一部分难治性靶基因的RNAi效率。这些结果表明，增强dsRNA稳定性不足以提高ECB中的RNAi效率，并表明存在导致ECB中低RNAi效率的额外的复杂机制。
Targeting insect-specific genes through post-transcriptional gene silencing with RNA interference (RNAi) is a new strategy for insect pest management. However, lepidopterans are recalcitrant to RNAi, which prevents the application of novel RNAi technology to many notorious pests, including Ostrinia nubilalis (ECB). Strategies for enhancing RNAi efficiency, including large doses of double-stranded RNA (dsRNA), nuclease inhibitors, transfection reagents, and nanoparticles, have proved useful in other insects exhibiting substantial dsRNA degradation, a major mechanism limiting RNAi efficacy. To determine if similar strategies can enhance RNAi efficiency in ECB, various reagents were tested for their ability to enhance dsRNA stability in ECB tissues, then compared for their effectiveness in whole ECB. Ex vivo incubation experiments revealed that Metafectene Pro, EDTA, chitosan-based dsRNA nanoparticles, and Zn2+ enhanced dsRNA stability in ECB hemolymph and gut content extracts, compared to uncoated dsRNA. Despite these positive results, the reagents used in this study were ineffective at enhancing RNAi efficiency in ECB in vivo. To reduce assay time and required dsRNA, midguts were dissected and incubated in tissue culture medium containing dsRNA with and without reagents. These experiments showed that RNAi efficiency varied between target genes, and nuclease inhibitors improved RNAi efficiency for only a portion of the refractory target genes investigated ex vivo. These results indicate that enhancing dsRNA stability is insufficient to improve RNAi efficiency in ECB and suggests the existence of additional, complex mechanisms contributing to low RNAi efficiency in ECB.
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影响因子：
3.7
作者：
Khajuria C;Ivashuta S;Wiggins E;Flagel L;Moar W;Pleau M;Miller K;Zhang Y;Ramaseshadri P;Jiang C;Hodge T;Jensen P;Chen M;Gowda A;McNulty B;Vazquez C;Bolognesi R;Haas J;Head G;Clark T
通讯作者：
Clark T
影响因子：
3.8
作者：
Arimatsu, Yuji;Kotani, Eiji;Furusawa, Toshiharu
通讯作者：
Furusawa, Toshiharu
影响因子：
2.2
作者：
Garbutt, Jennie S.;Belles, Xavier;Reynolds, Stuart E.
通讯作者：
Reynolds, Stuart E.
影响因子：
3.4
作者：
Park, Youngjin;Kim, Yonggyun
通讯作者：
Kim, Yonggyun
影响因子：
4.1
作者：
Castellanos, Nathaly L.;Smagghe, Guy;Christiaens, Olivier
通讯作者：
Christiaens, Olivier