Significantly Improved Recovery of Recombinant Sonchus Yellow Net Rhabdovirus by Expressing the Negative-Strand Genomic RNA
1
State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China
2
Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insect Pests, Zhejiang University, Hangzhou 310058, China
3
Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, Zhejiang University, Hangzhou 310058, China
*
Author to whom correspondence should be addressed.
Academic Editor: Henryk Czosnek
Viruses 2020, 12(12), 1459; https://doi.org/10.3390/v12121459
Received: 23 November 2020
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Revised: 10 December 2020
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Accepted: 15 December 2020
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Published: 17 December 2020
(This article belongs to the Section Viruses of Plants, Fungi and Protozoa)
Generation of recombinant negative-stranded RNA viruses (NSVs) from plasmids involves in vivo reconstitution of biologically active nucleocapsids and faces a unique antisense problem where the negative-sense viral genomic RNAs can hybridize to viral messenger RNAs. To overcome this problem, a positive-sense RNA approach has been devised through expression of viral antigenomic (ag)RNA and core proteins for assembly of antigenomic nucleocapsids. Although this detour strategy works for many NSVs, the process is still inefficient. Using Sonchus yellow net rhabdovirus (SYNV) as a model; here, we develop a negative-sense genomic RNA-based approach that increased rescue efficiency by two orders of magnitude compared to the conventional agRNA approach. The system relied on suppression of double-stranded RNA induced antiviral responses by co-expression of plant viruses-encoded RNA silencing suppressors or animal viruses-encoded double-stranded RNA antagonists. With the improved approach, we were able to recover a highly attenuated SYNV mutant with a deletion in the matrix protein gene which otherwise could not be rescued via the agRNA approach. Reverse genetics analyses of the generated mutant virus provided insights into SYNV virion assembly and morphogenesis. This approach may potentially be applicable to other NSVs of plants or animals.
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Keywords:
negative-stranded RNA virus; plant rhabdovirus; Sonchus yellow net virus; recovery; infectious clone; viral suppressor of RNA silencing; genome; antigenome; nucleocapsid; matrix protein
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MDPI and ACS Style
Ma, X.; Li, Z. Significantly Improved Recovery of Recombinant Sonchus Yellow Net Rhabdovirus by Expressing the Negative-Strand Genomic RNA. Viruses 2020, 12, 1459. https://doi.org/10.3390/v12121459
AMA Style
Ma X, Li Z. Significantly Improved Recovery of Recombinant Sonchus Yellow Net Rhabdovirus by Expressing the Negative-Strand Genomic RNA. Viruses. 2020; 12(12):1459. https://doi.org/10.3390/v12121459
Chicago/Turabian StyleMa, Xiaonan, and Zhenghe Li. 2020. "Significantly Improved Recovery of Recombinant Sonchus Yellow Net Rhabdovirus by Expressing the Negative-Strand Genomic RNA" Viruses 12, no. 12: 1459. https://doi.org/10.3390/v12121459
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