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化学修饰的双链 RNA 在昆虫的体外和体内诱导 RNAi 效应：一种提高基于 RNA 的植物保护的潜在新工具。

Chemically modified dsRNA induces RNAi effects in insects in vitro and in vivo: A potential new tool for improving RNA-based plant protection.

机构信息

Department of Chemical & Biological Engineering, University of Sheffield, Sheffield, United Kingdom.

Syngenta, Ghent Innovation Center, Ghent, Belgium.

出版信息

J Biol Chem. 2022 Sep;298(9):102311. doi: 10.1016/j.jbc.2022.102311. Epub 2022 Jul 31.

DOI:10.1016/j.jbc.2022.102311
PMID:35921898
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9478931/
Abstract

Global agriculture loses over $100 billion of produce annually to crop pests such as insects. Many of these crop pests either are not currently controlled by artificial means or have developed resistance against chemical pesticides. Long dsRNAs are capable of inducing RNAi in insects and are emerging as novel, highly selective alternatives for sustainable insect management strategies. However, there are significant challenges associated with RNAi efficacy in insects. In this study, we synthesized a range of chemically modified long dsRNAs in an approach to improve nuclease resistance and RNAi efficacy in insects. Our results showed that dsRNAs containing phosphorothioate modifications demonstrated increased resistance to southern green stink bug saliva nucleases. Phosphorothioate-modified and 2'-fluoro-modified dsRNA also demonstrated increased resistance to degradation by soil nucleases and increased RNAi efficacy in Drosophila melanogaster cell cultures. In live insects, we found chemically modified long dsRNAs successfully resulted in mortality in both stink bug and corn rootworm. These results provide further mechanistic insight into the dependence of RNAi efficacy on nucleotide modifications in the sense or antisense strand of the dsRNA in insects and demonstrate for the first time that RNAi can successfully be triggered by chemically modified long dsRNAs in insect cells or live insects.

摘要

全球农业每年因作物害虫（如昆虫）损失超过 1000 亿美元的农产品。许多这些作物害虫目前要么无法通过人工手段控制，要么已经对化学农药产生了抗药性。长 dsRNA 能够在昆虫中诱导 RNAi，并作为可持续昆虫管理策略的新型、高度选择性替代物而出现。然而，RNAi 在昆虫中的功效存在重大挑战。在这项研究中，我们通过合成一系列化学修饰的长 dsRNA，来提高昆虫中的核酸酶抗性和 RNAi 功效。我们的结果表明，含有硫代磷酸酯修饰的 dsRNA 显示出对南方绿臭虫唾液核酸酶的增强抗性。硫代磷酸酯修饰和 2'-氟修饰的 dsRNA 也显示出对土壤核酸酶降解的增强抗性，并在 Drosophila melanogaster 细胞培养物中提高了 RNAi 功效。在活体昆虫中，我们发现化学修饰的长 dsRNA 成功导致臭虫和玉米根虫的死亡。这些结果进一步深入了解了 RNAi 功效在昆虫 dsRNA 的正义或反义链上依赖核苷酸修饰的机制，并首次证明 RNAi 可以通过昆虫细胞或活体昆虫中的化学修饰的长 dsRNA 成功触发。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/91af2af0218d/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/dcab2975fbd6/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/d89deda8a614/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/6514b839a86b/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/d096ca228be9/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/d51e0db77b13/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/952c4c171bbb/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/ff45f9d3baf2/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/91af2af0218d/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/dcab2975fbd6/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/d89deda8a614/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/6514b839a86b/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/d096ca228be9/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/d51e0db77b13/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/952c4c171bbb/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/ff45f9d3baf2/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78df/9478931/91af2af0218d/gr8.jpg

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