Identification of plant RNA Editing factors in EMS mutants

本项目旨在通过筛选拟南芥EMS突变群体，通过靶向遗传方法进一步鉴定植物线粒体中RNA编辑的核编码因子。通过SNaPshot突变体筛选成功鉴定了几个新的PPR基因和新的MORF蛋白家族，证明了该方法提供了一个很好的工具。这项研究包括了迄今为止未知的酶和PPR的其他基因以及参与RNA编辑的其他蛋白质。已知PPR蛋白中的新点突变将允许阐明单个PPR蛋白结构域的功能。与此同时，线粒体编码蛋白的功能也将被探讨。位点特异性编辑蛋白将用于产生具有线粒体蛋白特异性改变的双敲除和amiRNA敲除突变体，以测定其功能，例如内含子matR阅读框。

This project aims to identify further nuclear encoded factors of RNA editing in mitochondria of plants through the targeted genetic approach by screening EMS mutant populations of Arabidopsis thaliana. The successful identification of several new PPR genes and the novel family of MORF proteins by the SNaPshot mutant screen proves that this approach provides an excellent tool. The search includes the so far unknown enzyme and further genes for PPR and other proteins involved in RNA editing. Novel point mutations in known PPR proteins will allow to elucidate the functions of individual PPR protein domains. In parallel, functions of mitochondrially encoded proteins will be probed. Site-specific editing proteins will be employed to generate double knock-out and amiRNA knock-down mutants with specific alterations in mitochondrial proteins to assay their function, for example of the intronic matR reading frame.