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A cytometry microparticle platform approach for screening tobacco microRNA changes after agrobacterium delivery

机译：一种用于农杆菌输送后筛选烟草microRNA变化的细胞计数微粒平台方法

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MicroRNAs are a class of non-coding regulatory RNAs that can modulate development as well as alter innate antiviral defenses in plants. In this study we explored changes in Nicotiana benthamiana tobacco microRNA expression as it relates to expression of a recombinant anti-Ebola GP1 antibody. The antibody was delivered to tobacco leaves through a bacterial Agrobacterium tumefaciens "agroinfiltration" expression strategy. A multiplex micro particle-based cytometry assay tracked the expression changes of 53 host tobacco microRNAs. Our results revealed that the most abundant microRNAs in actively growing leaves corresponded to nanoparticle probes specific to nta-mir-6149 and nta-miR-168b. After agroinfiltration, probes specific for nta-mir-398, and nta-mir-482d were significantly altered in their respective expression levels, however changes were partially attributed to the infiltration broth medium used in the antibody delivery process. Confirmation of nta-mir-398 and nta-mir-482d expression changes was also verified through RT-qPCR. To our knowledge this study is the first to profile medium and Agrobacterium injection at the microRNA level through a multiplex microparticle approach. (C) 2016 Elsevier B.V. All rights reserved.
机译：MicroRNA是一类非编码调控RNA，可以调节植物的发育以及改变其固有的抗病毒防御能力。在这项研究中，我们探讨了本氏烟草烟草microRNA表达的变化，因为它与重组抗埃博拉GP1抗体的表达有关。通过细菌根癌农杆菌“农杆菌浸润”表达策略将抗体递送至烟叶。基于多重微粒的细胞计数分析跟踪了53种宿主烟草microRNA的表达变化。我们的结果表明，活跃生长的叶片中最丰富的microRNA对应于nta-mir-6149和nta-miR-168b特异的纳米颗粒探针。农杆菌浸润后，对nta-mir-398和nta-mir-482d特异的探针在其各自的表达水平上均发生了显着变化，但是这种变化部分归因于抗体递送过程中使用的浸润肉汤培养基。还通过RT-qPCR验证了nta-mir-398和nta-mir-482d表达变化的确认。就我们所知，这项研究是第一个通过多重微粒方法在microRNA水平分析培养基和农杆菌注射的方法。 （C）2016 Elsevier B.V.保留所有权利。

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