Significantly Improved Recovery of Recombinant Sonchus Yellow Net Rhabdovirus by Expressing the Negative-Strand Genomic RNA
Autor: | Xiaonan Ma, Zhenghe Li |
---|---|
Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
0106 biological sciences
0301 basic medicine Gene Expression Regulation Viral viral suppressor of RNA silencing Transcription Genetic viruses Mutant negative-stranded RNA virus lcsh:QR1-502 nucleocapsid matrix protein Genome Viral Biology 01 natural sciences Virus Article lcsh:Microbiology Plant Viruses 03 medical and health sciences Sonchus recovery antigenome Genes Reporter Virology plant rhabdovirus Gene genome Plant Diseases Viral matrix protein Sonchus yellow net virus infectious clone RNA Reverse genetics Cell biology RNA silencing 030104 developmental biology Infectious Diseases Virion assembly RNA Viral RNA Interference Rhabdoviridae Genetic Engineering 010606 plant biology & botany |
Zdroj: | Viruses Volume 12 Issue 12 Viruses, Vol 12, Iss 1459, p 1459 (2020) |
ISSN: | 1999-4915 |
DOI: | 10.3390/v12121459 |
Popis: | 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. |
Databáze: | OpenAIRE |
Externí odkaz: | |
Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |