Abstract
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.
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