Rescue of Infectious Sindbis Virus by Yeast Spheroplast-Mammalian Cell Fusion.
Viruses
; 13(4)2021 04 01.
Article
in English
| MEDLINE | ID: covidwho-1389534
ABSTRACT
Sindbis virus (SINV), a positive-sense single stranded RNA virus that causes mild symptoms in humans, is transmitted by mosquito bites. SINV reverse genetics have many implications, not only in understanding alphavirus transmission, replication cycle, and virus-host interactions, but also in biotechnology and biomedical applications. The rescue of SINV infectious particles is usually achieved by transfecting susceptible cells (BHK-21) with SINV-infectious mRNA genomes generated from cDNA constructed via in vitro translation (IVT). That procedure is time consuming, costly, and relies heavily on reagent quality. Here, we constructed a novel infectious SINV cDNA construct that expresses its genomic RNA in yeast cells controlled by galactose induction. Using spheroplasts made from this yeast, we established a robust polyethylene glycol-mediated yeast BHK-21 fusion protocol to rescue infectious SINV particles. Our approach is timesaving and utilizes common lab reagents for SINV rescue. It could be a useful tool for the rescue of large single strand RNA viruses, such as SARS-CoV-2.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Sindbis Virus
/
Spheroplasts
/
Yeasts
/
Cell Fusion
/
Alphavirus Infections
/
Host Microbial Interactions
Type of study:
Experimental Studies
Limits:
Animals
Language:
English
Year:
2021
Document Type:
Article
Affiliation country:
V13040603
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