Department of Biological Sciences, Mississippi State University, Mississippi State, MS, USA.
Methods Mol Biol. 2022;2316:29-37. doi: 10.1007/978-1-0716-1464-8_3.
Systemic RNA trafficking widely exists in plants and is critical for integrating the healthy development and responses to environmental cues. Viroids, single-stranded circular noncoding RNAs that infect plants, have been used as a model to delineate the mechanism underlying systemic RNA trafficking. Recent work on viroids has shown that structural motifs are critical to direct RNA trafficking through distinct cellular boundaries. Here, we describe the methods for generating mutational variants using site-directed mutagenesis and infection assays to unravel the function of RNA motifs. This approach can be modified to study other RNA motif-based biological processes.
系统 RNA 运输广泛存在于植物中,对于整合健康发育和对环境线索的反应至关重要。感染植物的单链环状非编码 RNA 类病毒已被用作模型来描绘系统 RNA 运输的机制。最近对类病毒的研究表明,结构基序对于通过不同的细胞边界直接进行 RNA 运输至关重要。在这里,我们描述了使用定点诱变和感染测定来生成突变变体的方法,以阐明 RNA 基序的功能。这种方法可以修改用于研究其他基于 RNA 基序的生物学过程。