病毒诱导基因沉默(VIGS)在植物功能基因组学研究中的应用

Dagnachew Bekele, K. Tesfaye, A. Fikre
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引用次数: 14

摘要

病毒诱导基因沉默(VIGS)是一种利用植物抗病毒防御机制的有效技术。基因转录抑制技术是近年来发展起来的一种研究植物基因功能的技术。VIGS是一种快速、高效、特异的瞬时基因沉默系统。VIGS的主要步骤包括:将病毒基因组改造成合适的病毒载体,将目标被沉默的宿主基因片段整合进去,感染合适的植物宿主,并使目标基因沉默,作为植物抵御病毒感染的防御机制的一部分。VIGS载体是一种重组病毒,可以携带来自宿主的内源性基因片段。在被修饰载体感染的过程中,宿主会对克隆的宿主基因产生防御反应;功能表型的缺失使得鉴定基因的功能成为可能。重组病毒通过农杆菌介导的瞬时表达或体外转录RNA接种或直接DNA接种导入植物细胞。Trans基因与病毒RNA一起被RNA依赖的RNA聚合酶扩增,产生dsRNA分子。dsRNA是转录后基因沉默的触发分子。作为功能基因组学研究的反向遗传学工具,VIGS具有许多优势。尽管潜力巨大,但仍有许多限制有待克服。本文就VIGS技术的分子机制、在植物功能基因组学研究中的最新应用、主要局限及未来发展前景进行了综述。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Applications of Virus Induced Gene Silencing (VIGS) in Plant Functional Genomics Studies
Virus induced gene silencing (VIGS) is an effective technology that exploits an antiviral defense mechanism in plants. It is a recently developed gene transcript suppression technique for characterizing the function of plant genes. VIGS is rapid, efficient and specific system for transient gene silencing. The major steps in VIGS includes; engineering viral genomes to the appropriate viral vector to incorporate fragments of host genes that are targeted to be silenced, infecting the appropriate plant hosts and silencing the target genes as part of the defense mechanism of the plant against virus infection. The VIGS vector is a recombinant virus engineered to be able to carry a piece an endogenous gene from the host. During infection with the modified vector, the host’s defense reaction will be induced against the cloned host gene; a loss of function phenotype makes it possible to identify the function of the gene. The recombinant virus is introduced into plant cells through Agrobacterium tumefaciens mediated transient expression or in vitro transcribed RNA inoculation or direct DNA inoculation. The Trans gene is amplified along with the viral RNA by RNA dependent RNA polymerase generating dsRNA molecules. dsRNA is the triggering molecule of Post transcriptional gene silencing. VIGS as a reverse genetics tool for functional genomics studies presenting several advantages. Despite its great potential, many limitations remain to be overcome. In this review, the molecular mechanism in VIGS technology, its advanced application in plant functional genomics studies and the major limitation and potential future prospects were briefly discussed.
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