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VipariNama: RNA vectors to rapidly reprogram plant morphology and metabolism
bioRxiv - Plant Biology Pub Date : 2020-06-04 , DOI: 10.1101/2020.06.03.130179
Arjun Khakhar , Cecily Wang , Ryan Swanson , Sydney Stokke , Furva Rizvi , Surbhi Sarup , John Hobbs , Daniel F. Voytas

Synthetic transcription factors have great promise as tools to explore biological processes. By allowing precise alterations in gene expression, they can help elucidate relationships between gene expression and plant morphology or metabolism. However, the years-long timescales, high cost, and technical skill associated with plant transformation have dramatically slowed their use. In this work, we developed a new platform technology called VipariNama (ViN) in which RNA vectors are used to rapidly deploy synthetic transcription factors and reprogram gene expression in planta. We demonstrate how ViN vectors can direct activation or repression of multiple genes, systemically and persistently over several weeks, and in multiple plant species. We also show how this transcriptional reprogramming can create predictable changes to metabolic and morphological phenotypes in the model plants Nicotiana benthamiana and Arabidopsis thaliana in a matter of weeks. Finally, we show how a model of gibberellin signaling can guide ViN vector-based reprogramming to rapidly engineer plant size in both model species as well as the crop Solanum lycopersicum (tomato). In summary, using VipariNama accelerates the timeline for generating phenotypes from over a year to just a few weeks, providing an attractive alternative to transgenesis for synthetic transcription factor-enabled hypothesis testing and crop engineering.

中文翻译:

VipariNama:RNA载体可快速重编程植物形态和代谢

合成转录因子有望成为探索生物学过程的工具。通过允许基因表达的精确改变,它们可以帮助阐明基因表达与植物形态或代谢之间的关系。然而,长达数年的时间尺度,高成本以及与植物转化相关的技术技能极大地减缓了它们的使用。在这项工作中,我们开发了一种称为VipariNama(ViN)的新平台技术,其中RNA载体用于在植物中快速部署合成转录因子并重新编程基因表达。我们展示了ViN载体如何在几个星期内以及在多个植物物种中系统地,持续地指导多个基因的激活或抑制。我们还展示了这种转录重编程如何在数周内在模型植物烟草本氏烟草和拟南芥中产生代谢和形态表型的可预测变化。最后,我们展示了赤霉素信号传导模型如何指导基于ViN载体的重编程,以快速改造模型物种以及农作物茄子(番茄)中的植物大小。总之,使用VipariNama可以将生成表型的时间表从一年以上缩短到仅仅几周,为转基因技术提供了有吸引力的替代方法,用于合成转录因子假设研究和作物工程。我们展示了赤霉素信号传导模型如何指导基于ViN载体的重编程,以快速设计模型物种以及农作物茄子(番茄)中的植物大小。总之,使用VipariNama可以将生成表型的时间表从一年以上缩短到仅仅几周,为转基因技术提供了有吸引力的替代方法,用于合成转录因子假设研究和作物工程。我们展示了赤霉素信号传导模型如何指导基于ViN载体的重编程,以快速设计模型物种以及农作物茄子(番茄)中的植物大小。总之,使用VipariNama可以将生成表型的时间表从一年以上缩短到仅仅几周,为转基因技术提供了有吸引力的替代方法,用于合成转录因子假设研究和作物工程。
更新日期:2020-06-04
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