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基于 RNAi 的基因沉默策略在植物抗病毒病中的应用。

Strategies for Efficient RNAi-Based Gene Silencing of Viral Genes for Disease Resistance in Plants.

机构信息

Department of Plant Biotechnology, Centre for Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India.

出版信息

Methods Mol Biol. 2022;2408:23-35. doi: 10.1007/978-1-0716-1875-2_2.

Abstract

RNA interference (RNAi) is an evolutionarily conserved gene silencing mechanism in eukaryotes including fungi, plants, and animals. In plants, gene silencing regulates gene expression, provides genome stability, and protect against invading viruses. During plant virus interaction, viral genome derived siRNAs (vsiRNA) are produced to mediate gene silencing of viral genes to prevent virus multiplication. After the discovery of RNAi phenomenon in eukaryotes, it is used as a powerful tool to engineer plant viral disease resistance against both RNA and DNA viruses. Despite several successful reports on employing RNA silencing methods to engineer plant for viral disease resistance, only a few of them have reached the commercial stage owing to lack of complete protection against the intended virus. Based on the knowledge accumulated over the years on genetic engineering for viral disease resistance, there is scope for effective viral disease control through careful design of RNAi gene construct. The selection of target viral gene(s) for developing the hairpin RNAi (hp-RNAi) construct is very critical for effective protection against the viral disease. Different approaches and bioinformatics tools which can be employed for effective target selection are discussed. The selection of suitable target regions for RNAi vector construction can help to achieve a high level of transgenic virus resistance.

摘要

RNA 干扰 (RNAi) 是真核生物(包括真菌、植物和动物)中一种进化上保守的基因沉默机制。在植物中,基因沉默调节基因表达、提供基因组稳定性并抵御入侵病毒。在植物病毒相互作用过程中,产生了源自病毒基因组的 siRNA(vsiRNA),以介导病毒基因的基因沉默,从而阻止病毒复制。在真核生物中发现 RNAi 现象后,它被用作一种强大的工具,用于工程植物对 RNA 和 DNA 病毒的抗病毒病能力。尽管有几项关于利用 RNA 沉默方法来工程植物抗病毒病的成功报告,但由于缺乏对预期病毒的完全保护,只有少数报告进入了商业阶段。基于多年来在抗病毒病遗传工程方面积累的知识,通过精心设计 RNAi 基因构建,可以有效地控制病毒病。开发发夹 RNAi (hp-RNAi) 构建体时,选择目标病毒基因 (s) 非常关键,这对于有效保护病毒病至关重要。讨论了可用于有效目标选择的不同方法和生物信息学工具。选择用于 RNAi 载体构建的合适靶区有助于实现高水平的转基因病毒抗性。

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