Identifying genes involved in microRNA turnover in Arabidopsis

鉴定拟南芥中参与 microRNA 周转的基因

• 批准号: 1799918
• 金额: --
• 依托单位: University of East Anglia
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1799918
• 关键词: Identifying; genes; involved; microRNA; turnover

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 that 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 successful candidate 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 will be induced by germinating seeds on a special media. After changing the media the miRNA transcription is switched off and the existing miRNA molecules will be degraded, leading to green fluorescence recovery in the seedlings. The successful candidate will screen for lines exhibiting faster or slower GFP recovery. After validation of the mutant phenotype the student will identify the mutant genes 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 aspect of plant biology but also on biomedicine as siRNAs are potential therapeutics.

microRNA（miRNAs）是一类非编码RNA小分子，通过靶向mRNA调控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响应中起着关键作用，因此miRNA在正确的时间和地点表达是很重要的。因此，必须有一种降解miRNA的机制（miRNA周转），并且必须仔细调节该机制本身，以确保在不需要其功能时不存在miRNA。然而，我们对这种机制及其调控知之甚少。成功的候选人将进行突变体筛选，以确定参与miRNA周转的基因。筛选测定依赖于GFP传感器，其被miRNA靶向。通过在特殊培养基上萌发种子来诱导miRNA的表达。更换培养基后，miRNA转录被关闭，现有的miRNA分子将被降解，导致幼苗中的绿色荧光恢复。成功的候选者将筛选表现出更快或更慢GFP恢复的细胞系。在突变表型验证后，学生将通过下一代测序来识别突变基因。识别参与miRNA周转的基因将为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周转如何调节将影响植物生物学的各个方面，也会影响生物医学，因为siRNA是潜在的治疗方法。