Chapter title |
Transiently Induce RNA Silencing in Plants Using a Tobacco Necrosis Virus A (TNV-A)-Based dsRNA Production System.
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Book title |
Double-Stranded RNA
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Published in |
Methods in molecular biology, January 2024
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DOI | 10.1007/978-1-0716-3702-9_12 |
Pubmed ID | |
Book ISBNs |
978-1-07-163701-2, 978-1-07-163702-9
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Authors |
Zhang, Yuanming, Chai, Mengzhu, Cheng, Xiaofei, Xu, Kai |
Abstract |
Transgenic expression of hairpin RNA or artificial microRNA is widely used for genetic studies in plant science. However, induction of RNA silencing by transgenic method may have a problem when studying essential genes. Here, we provide an in planta transient double-stranded RNA (dsRNA) producing system using a tobacco necrosis virus A (TNV-A)-based replicon for efficiently inducing RNA silencing in plants. In this system, the target sequence is placed between the cauliflower mosaic virus 35S promoter and the 3'-terminal part of viral genomic RNA, while the C-terminal part of TNV-A RNA-dependent RNA polymerase (p82C) is expressed by a different promoter. The endogenous RNA polymerase-synthesized target sequence is recruited by p82C to produce dsRNA to induce RNA silencing. |
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