Characterising genes involved in microRNA degradation in Arabidopsis

MicroRNAs(MiRNAs)是一种小的非编码RNA分子，通过靶向mRNAs来调节蛋白质编码基因的表达。由于它们在发育以及对生物和非生物胁迫的反应中发挥关键作用，因此在正确的时间和地点表达miRNAs是很重要的。因此，必须有一种机制来降解miRNAs(miRNA周转)，并且这种机制本身必须受到仔细的监管，以确保当miRNAs的功能不需要时，miRNAs不存在。然而，我们对这一机制及其调控知之甚少。该项目团队将进行突变筛选，以确定与miRNA周转有关的基因。筛选试验依赖于GFP传感器，该传感器以miRNA为靶标。MiRNA的表达是通过在特殊的培养基上萌发种子来诱导的。更换介质后，miRNA转录被关闭，现有的miRNA分子被降解，导致幼苗中绿色荧光的恢复。该项目将筛选出GFP恢复较快或较慢的线路。在确认突变表型后，将通过下一代测序来鉴定突变基因。识别参与miRNA周转的基因将在RNA沉默领域开辟一个新的领域。了解这些基因的功能以及miRNA周转是如何调节的，不仅将影响植物生物学的各个方面，还将影响生物医学，因为siRNA是潜在的治疗方法。该项目将在一个充满活力和活力的研究小组中进行，成员包括湿法实验室研究人员和生物信息学家。

MicroRNAs (miRNAs) are small non-coding RNA molecules regulating the expression of protein coding genes by targeting mRNAs. Since they play key roles in development and in responses to both biotic and abiotic stresses, it is important that miRNAs are expressed at the right time and place. Therefore there must be a mechanism to degrade miRNAs (miRNA turnover) and this mechanism itself must be carefully regulated to ensure that miRNAs are not present when their function is not required. However, we know very little about this mechanism and its regulation. The project team will carry out a mutant screen to identify genes involved in miRNA turnover. The screening assay relies on a GFP sensor, which is targeted by a miRNA. Expression of the miRNA is induced by germinating seeds on a special media. After changing the media the miRNA transcription is switched off and the existing miRNA molecules are degraded, leading to green fluorescence recovery in the seedlings. The project will screen for lines exhibiting faster or slower GFP recovery. After validation of the mutant phenotype the mutant genes will be identified by next generation sequencing. Identifying genes involved in miRNA turnover will open up a new field in the RNA silencing area. Understanding the function of those genes and how miRNA turnover is regulated will impact on all aspects of plant biology but also on biomedicine as siRNAs are potential therapeutics. The project will run in a vibrant and dynamic research group including wet lab researchers and bioinformaticians.