Open AccessPoliovirus Replicon RNA Generation, Transfection, Packaging, and Quantitation of Replication
Author(s) -
Viktorova Ekaterina G.,
Khattar Sunil,
Samal Siba,
Belov George A.
Publication year - 2018
Publication title -
current protocols in microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.813
H-Index - 35
eISSN - 1934-8533
pISSN - 1934-8525
DOI - 10.1002/cpmc.47
Subject(s) - replicon , biology , poliovirus , rna , picornaviridae , virology , luciferase , viral replication , transfection , reporter gene , virus , microbiology and biotechnology , gene , genetics , genome , gene expression
Poliovirus is a prototype member of the Enterovirus genus of the Picornaviridae family of small positive strand RNA viruses, which include important human and animal pathogens. Quantitative assessment of viral replication is very important for investigation of the virus biology and the development of anti‐viral strategies. The poliovirus genome structure allows replacement of structural genes with a reporter protein, such as a luciferase or a fluorescent protein, whose signals can be detected and quantified in vivo , thus permitting observation of replication kinetics in live cells. This paper presents protocols for poliovirus replicon RNA production, purification, packaging and transfection, as well as techniques for monitoring Renilla luciferase replication signal in living cells. © 2018 by John Wiley & Sons, Inc.