Open AccessMulti-color fluorescent reporter dengue viruses with improved stability for analysis of a multi-virus infection
Author(s) -
Amporn Suphatrakul,
Thaneeya Duangchinda,
Natapong Jupatanakul,
Kanjanawadee Prasittisa,
Suppachoke Onnome,
Jutharat Pengon,
Bunpote Siridechadilok
Publication year - 2018
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0194399
Subject(s) - biology , reporter gene , virology , dengue virus , virus , viral replication , green fluorescent protein , capsid , dengue fever , gene , genetics , gene expression
Reporter virus is a versatile tool to visualize and to analyze virus infections. However, for flaviviruses, it is difficult to maintain the inserted reporter genes on the viral genome, limiting its use in several studies that require homogeneous virus particles and several rounds of virus replication. Here, we showed that flanking inserted GFP genes on both sides with ribosome-skipping 2A sequences improved the stability and the consistency of their fluorescent signals for dengue-virus-serotype 2 (DENV2) reporter viruses. The reporter viruses can infect known susceptible mammalian cell lines and primary CD14+ human monocytes. This design can accommodate several fluorescent protein genes, enabling the generation of multi-color DENV2-16681 reporter viruses with comparable replication capabilities, as demonstrated by their abilities to maintain their fluorescent intensities during co-infections and to exclude superinfections regardless of the fluorescent tags. The reported design of multi-color DENV2 should be useful for high-throughput analyses, single-cell analysis, and characterizations of interference and superinfection in animal models.