Stanfield Alyssa M, May Jared P
Division of Biological & Biomedical Systems, School of Science and Engineering, University of Missouri-Kansas City, Kansas City, Missouri, USA.
Mol Plant Pathol. 2025 Jun;26(6):e70113. doi: 10.1111/mpp.70113.
Artificial condensates formed by liquid-liquid phase separation (LLPS) have been extensively studied in the field of synthetic biology for their ability to control cellular functions in a reversible and tunable manner. Although cellular condensates can restrict viral accumulation, the potential to employ LLPS as a platform for targeted antiviral strategies remains understudied. Here, we engineered tobacco mosaic virus (TMV) to harbour MS2 bacteriophage hairpins that are specifically bound by the MS2 coat protein (MCP). Using well-studied intrinsically disordered regions (IDRs) as scaffolds, we engineered artificial condensates that could target MS2-tagged TMV, inhibiting TMV accumulation up to fivefold following transient expression in Nicotiana benthamiana. TAR DNA-binding protein 43 (TDP-43)-based condensates inhibited MS2-tagged virus accumulation by over twofold, independent of the global translational repression observed during condensate expression. Following manual inoculation of MS2-tagged TMV onto leaves transiently expressing TDP-43:MCP condensates, systemic virus accumulation was reduced by over 10-fold, which was associated with a significant decrease in symptom severity. These results provide a foundation for developing artificial biomolecular condensates to mitigate plant virus accumulation and disease severity.
通过液-液相分离（LLPS）形成的人工凝聚物因其能够以可逆和可调的方式控制细胞功能而在合成生物学领域得到了广泛研究。尽管细胞凝聚物可以限制病毒积累，但将LLPS用作靶向抗病毒策略平台的潜力仍未得到充分研究。在这里，我们对烟草花叶病毒（TMV）进行工程改造，使其携带能被MS2外壳蛋白（MCP）特异性结合的MS2噬菌体发夹结构。利用经过充分研究的内在无序区域（IDR）作为支架，我们构建了能够靶向MS2标记的TMV的人工凝聚物，在本氏烟草中瞬时表达后，可将TMV积累抑制高达五倍。基于TAR DNA结合蛋白43（TDP-43）的凝聚物将MS2标记的病毒积累抑制了两倍多，这与凝聚物表达过程中观察到的全局翻译抑制无关。在将MS2标记的TMV人工接种到瞬时表达TDP-43:MCP凝聚物的叶片上后，系统病毒积累减少了10倍以上，这与症状严重程度的显著降低有关。这些结果为开发人工生物分子凝聚物以减轻植物病毒积累和疾病严重程度奠定了基础。
