Avital Aviram, Muzika Noy Sadot, Persky Zohar, Karny Avishai, Bar Gili, Michaeli Yuval, Shklover Jeny, Shainsky Janna, Weissman Haim, Shoseyov Oded, Schroeder Avi
Laboratory for Targeted Drug Delivery and Personalized Medicine Technologies, Department of Chemical Engineering, Technion - Israel Institute of Technology, Haifa 3200003, Israel.
The Norman Seiden Multidisciplinary Program for Nanoscience and Nanotechnology, Technion - Israel Institute of Technology, Haifa 3200003, Israel.
Adv Funct Mater. 2021 Jul 10;31(44). doi: 10.1002/adfm.202101003. eCollection 2021 Oct 26.
Grapevine leafroll disease (GLD) is a globally spreading viral infection that causes major economic losses by reducing crop yield, plant longevity and berry quality, with no effective treatment. Grapevine leafroll associated virus-3 (GLRaV-3) is the most severe and prevalent GLD strain. Here, we evaluated the ability of RNA interference (RNAi), a non-GMO gene-silencing pathway, to treat GLRaV-3 in infected Cabernet Sauvignon grapevines. We synthesized lipid-modified polyethylenimine (lmPEI) as a carrier for long double-stranded RNA (dsRNA, 250-bp-long) that targets RNA polymerase and coat protein genes that are conserved in the GLRaV-3 genome. Self-assembled dsRNA-lmPEI particles, 220 nm in diameter, displayed inner ordered domains spaced 7.3±2 nm from one another, correlating to lmPEI wrapping spirally around the dsRNA. The particles effectively protected RNA from degradation by ribonucleases, and Europium-loaded particles applied to grapevine leaves were detected as far as 60-cm from the foliar application point. In three field experiments, a single dose of foliar administration knocked down GLRaV-3 titer, and multiple doses of the treatment kept the viral titer at baseline and triggered recovery of the vine and berries. This study demonstrates RNAi as a promising platform for treating viral diseases in agriculture.
葡萄卷叶病（GLD）是一种在全球范围内传播的病毒感染，它会降低作物产量、植株寿命和浆果品质，从而造成重大经济损失，且目前尚无有效治疗方法。葡萄卷叶相关病毒3型（GLRaV-3）是最严重且最普遍的GLD毒株。在此，我们评估了RNA干扰（RNAi）这种非转基因基因沉默途径在感染赤霞珠葡萄植株中治疗GLRaV-3的能力。我们合成了脂质修饰的聚乙烯亚胺（lmPEI）作为长双链RNA（dsRNA，250个碱基对长）的载体，该双链RNA靶向GLRaV-3基因组中保守的RNA聚合酶和外壳蛋白基因。自组装的直径为220 nm的dsRNA-lmPEI颗粒显示出内部有序结构域，彼此间隔7.3±2 nm，这与lmPEI围绕dsRNA螺旋缠绕相关。这些颗粒有效地保护RNA不被核糖核酸酶降解，并且在葡萄叶片上施用的负载铕的颗粒在距离叶面施药点60厘米处仍能被检测到。在三项田间试验中，单次叶面给药降低了GLRaV-3的滴度，多次给药使病毒滴度维持在基线水平，并促使葡萄藤和浆果恢复。这项研究证明RNAi是农业中治疗病毒病害的一个有前景的平台。
