工具与靶点:植物病毒在CRISPR-Cas基因组编辑中的双重作用
Tools and targets: The dual role of plant viruses in CRISPR-Cas genome editing.
作者信息
Uranga Mireia, Daròs José-Antonio
机构信息
Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas - University. Politècnica de València, Valencia, 46022, Spain.
出版信息
Plant Genome. 2023 Jun;16(2):e20220. doi: 10.1002/tpg2.20220. Epub 2022 Jun 14.
The recent emergence of tools based on the clustered, regularly interspaced, short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins have revolutionized targeted genome editing, thus holding great promise to both basic plant science and precision crop breeding. Conventional approaches for the delivery of editing components rely on transformation technologies or transient delivery to protoplasts, both of which are time-consuming, laborious, and can raise legal concerns. Alternatively, plant RNA viruses can be used as transient delivery vectors of CRISPR-Cas reaction components, following the so-called virus-induced genome editing (VIGE). During the last years, researchers have been able to engineer viral vectors for the delivery of CRISPR guide RNAs and Cas nucleases. Considering that each viral vector is limited to its molecular biology properties and a specific host range, here we review recent advances for improving the VIGE toolbox with a special focus on strategies to achieve tissue-culture-free editing in plants. We also explore the utility of CRISPR-Cas technology to enhance biotic resistance with a special focus on plant virus diseases. This can be achieved by either targeting the viral genome or modifying essential host susceptibility genes that mediate in the infection process. Finally, we discuss the challenges and potential that VIGE holds in future breeding technologies.
最近出现的基于成簇规律间隔短回文重复序列(CRISPR)和CRISPR相关(Cas)蛋白的工具彻底改变了靶向基因组编辑,因此对基础植物科学和精准作物育种都具有巨大的前景。传统的编辑组件递送方法依赖于转化技术或向原生质体的瞬时递送,这两种方法都既耗时又费力,还可能引发法律问题。另外,植物RNA病毒可以用作CRISPR-Cas反应组件的瞬时递送载体,即所谓的病毒诱导基因组编辑(VIGE)。在过去几年中,研究人员已经能够构建用于递送CRISPR引导RNA和Cas核酸酶的病毒载体。鉴于每种病毒载体都受其分子生物学特性和特定宿主范围的限制,在此我们综述了改进VIGE工具箱的最新进展,特别关注在植物中实现无组织培养编辑的策略。我们还探讨了CRISPR-Cas技术在增强生物抗性方面的应用,特别关注植物病毒病。这可以通过靶向病毒基因组或修饰在感染过程中起介导作用的关键宿主易感性基因来实现。最后,我们讨论了VIGE在未来育种技术中面临的挑战和潜力。