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确定植物线粒体中RNA编辑的核编码因子。通过快照突变筛选成功地鉴定了几个新的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。搜索包括迄今未知的酶和PPR的进一步基因，以及参与RNA编辑的其他蛋白质。已知的PPR蛋白的新的点突变将有助于阐明单个PPR蛋白结构域的功能。同时，还将探索线粒体编码蛋白的功能。定点编辑蛋白将被用来产生具有线粒体蛋白特定变化的双重敲除和amiRNA敲除突变体，以分析它们的功能，例如内含子读码框架。

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.