Grinding the infected N. benthamiana leaves and injecting the extracts into the thoraxes of SBPHs. 2015). Foxtail mosaic virus: a viral vector for protein expression in cereals. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, et al. Ahlquist P, French R, Janda M, Loesch-Fries LS. State Key Laboratory of Agro-Biotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China, Ying Zang, Xiao-Dong Fang, Ji-Hui Qiao, Qiang Gao & Xian-Bing Wang, You can also search for this author in Hum Genet. Bouton C, King RC, Chen H, Azhakanandam K, Bieri S, Hammond-Kosack KE, et al. Tomato spotted wilt virus is a segmented NSR virus and belongs to the genus Orthotospovirus in the family Tospoviridae (Kormelink et al. 2017;8:539. 2011;162:110–25. Plant Biotechnol J. The NSR virus genomes are complementary to their mRNA and have no infectious activity alone. Multiplex genome engineering using CRISPR/Cas systems. German TL, Lorenzen MD, Grubbs N, Whitfield AE. Brome mosaic virus (BMV) was the first plant virus to be rescued from cloned viral cDNA in 1984 (Ahlquist et al. Since then, reverse genetics systems of numerous plant positive-strand RNA viruses have been developed and exploited as vectors for gene expression and virus-induced RNA silencing (Hefferon 2014; Cody and Scholthof 2019). Sun K, Zhou X, Lin W, Zhou X, Jackson AO, Li Z. Matrix-glycoprotein interactions required for budding of a plant nucleorhabdovirus and induction of inner nuclear membrane invagination. 2015;8:1288–91. NLM 2008). Arch Virol. French R, Janda M, Ahlquist P. Bacterial gene inserted in an engineered RNA virus: efficient expression in monocotyledonous plant cells. In the middle panels, combined technologies were used for developing the reverse genetics systems of SYNV, BYSMV, TSWV, and RRV. FUTURE APPLICATIONS OF REVERSE GENETICS SYSTEMS. Virus Res. Mol Plant Pathol. Currently, some of these viral vectors have been popularly used to produce plant-derived biopharmaceuticals, such as monoclonal antibodies, vaccines and other therapeutic proteins (Rybicki 2009; Rybicki 2010; Saxena et al. 2019). In contrast, The RNA5, RNA6, and RNA7 are not essential for RRV infections (Di Bello et al. 1984 Nov;81(21):6812-6 Nature. e Codon optimization removed cryptic splicing sites in the cDNA of TSWV L and M RNAs for stable expression of the large RdRp protein and genome RNAs in plants. By using this website, you agree to our Annu Rev Phytopathol. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. 2020. As described above, plant rhabdoviruses could stably express multiple and large foreign proteins, which expends the research into expression of both the sgRNA and the Cas9 nuclease in one virus vector. Changes to virus taxonomy and the international code of virus classification and nomenclature ratified by the international committee on taxonomy of viruses. Curr Opin Biotech. 2020). PubMed Central  All authors read and approved the final manuscript. Classical plant rhabdoviruses are non-segmented NSR viruses and consist of the Cytorhabdovirus and Nucleorhabdovirus genera according to their replication and morphogenesis sites on cytoplasm or nucleus. 1385. 2020). 2019b;93:e00680-19. Yan T, Zhu JR, Di D, Gao Q, Zhang Y, Zhang A, et al. 2013). Reverse Genetics Systems for Negative-Sense RNA Viruses. 2015;210:241–4. Sci Rep. 2017;7:12647. 2011). Plant NSR viruses consist of members of the families Rhabdoviridae, Tospoviridae, Phenuiviridae, Aspiviridae, and Fimoviridae and the unclassified genus Coguvirus (Abudurexiti et al. Last and importantly, it should be emphasized that biosafety should be carefully considered before implementation of these plant virus vectors in fields. 2015). Two years later, the first plant RNA virus vector was developed based on BMV (French et al. Methods in molecular biology, vol. 2019). Then, these BYSMV-RFP-infected SBPHs were transferred to healthy barley plants. Biomedicines. The CCR4 proteins were recruited by P into the N-RNA complexes and induced turnover of cellular mRNAs that nonspecifically bind to the BYSMV N protein (Zhang et al. Therefore, these newly established reverse genetics systems of plant NSR viruses would allow dissection of detailed molecular mechanisms of virus-host interactions in the context of virus infection. Arch Virol. Consequently, the codon-optimized RdRp, rather than the wild-type RdRp protein, supported transcription and replication of the TSWV S gRNA-based MR system when co-expressed with N protein and four VSRs. 2020). Ishibashi K, Matsumoto-Yokoyama E, Ishikawa M. A tomato spotted wilt virus S RNA-based replicon system in yeast. Rhabdovirus genomes encode five structural proteins, including the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and the large protein of viral polymerase (L) (Jackson et al.

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