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Enhanced virus-mediated gene-editing in plant meristematic tissues

增强植物分生组织中病毒介导的基因编辑

基本信息

批准号：
1897792
负责人：
金额：
--
依托单位：
University of Warwick
依托单位国家：
英国
项目类别：
Studentship
财政年份：
2017
资助国家：
英国
起止时间：
2017 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=studentship-1897792
关键词：
Enhanced virus mediated gene editing

项目摘要

For many years it has been possible to manipulate aspects of plant development through the creation of transgenic plants expressing exogenous genes, however it has always been a long-term goal to do this by altering endogenous genes without the need for transgenic plants. Recently technologies have been developed that now enable gene-editing of endogenous genes, such techniques include Zinc finger nucleases, TALENs, and the more recent CRISPR RNA gene editing system. We will use the CRISPR system, to alter the sequences of genes involved in the control of flowering in order to manipulate the flowering time of crop plants. We have a virus-based expression system which we have already used successfully to express functional proteins in tobacco plants. We will test whether this expression system can be used to deliver the gene-editing proteins/RNA into plant cells and to create gene-edited plants. In this project we will address one of the main constraints of this approach which is to get the virus to express the Cas9 protein and sgRNAs in the shoot apical meristem so that gene editing changes can be made in germ-line cells to produce heritable changes. If such approaches are successful then this will revolutionize the way in which genetically modified plants are produced, enabling them to be produced without the need to incorporate any foreign DNA into their genome.

多年来，通过产生表达外源基因的转基因植物来操纵植物发育的各个方面是可能的，然而，通过改变内源基因而不需要转基因植物来实现这一点一直是长期目标。最近已经开发了能够对内源基因进行基因编辑的技术，这些技术包括锌指核酸酶、TALEN和最近的CRISPR RNA基因编辑系统。我们将使用CRISPR系统来改变控制开花的基因序列，以操纵作物的开花时间。我们有一个基于病毒的表达系统，我们已经成功地用于在烟草植物中表达功能蛋白。我们将测试这种表达系统是否可用于将基因编辑蛋白/RNA递送到植物细胞中并创建基因编辑植物。在这个项目中，我们将解决这种方法的主要限制之一，即让病毒在茎尖分生组织中表达Cas9蛋白和sgRNA，以便在生殖系细胞中进行基因编辑改变，从而产生可遗传的变化。如果这些方法成功，那么这将彻底改变转基因植物的生产方式，使它们能够在不需要将任何外源DNA掺入其基因组的情况下生产。