TY - JOUR
T1 - An Improved Reverse Genetics System to Overcome Cell-Type-Dependent Ebola Virus Genome Plasticity
AU - Tsuda, Yoshimi
AU - Hoenen, Thomas
AU - Banadyga, Logan
AU - Weisend, Carla
AU - Ricklefs, Stacy M.
AU - Porcella, Stephen F.
AU - Ebihara, Hideki
N1 - Publisher Copyright:
© 2015 Published by Oxford University Press on behalf of the Infectious Diseases Society of America 2015. This work is written by (a) US Government employee(s) and is in the public domain in the US.
PY - 2015/10/1
Y1 - 2015/10/1
N2 - Reverse genetics systems represent a key technique for studying replication and pathogenesis of viruses, including Ebola virus (EBOV). During the rescue of recombinant EBOV from Vero cells, a high frequency of mutations was observed throughout the genomes of rescued viruses, including at the RNA editing site of the glycoprotein gene. The influence that such genomic instability could have on downstream uses of rescued virus may be detrimental, and we therefore sought to improve the rescue system. Here we report an improved EBOV rescue system with higher efficiency and genome stability, using a modified full-length EBOV clone in Huh7 cells. Moreover, by evaluating a variety of cells lines, we revealed that EBOV genome instability is cell-type dependent, a fact that has significant implications for the preparation of standard virus stocks. Thus, our improved rescue system will have an impact on both basic and translational research in the filovirus field.
AB - Reverse genetics systems represent a key technique for studying replication and pathogenesis of viruses, including Ebola virus (EBOV). During the rescue of recombinant EBOV from Vero cells, a high frequency of mutations was observed throughout the genomes of rescued viruses, including at the RNA editing site of the glycoprotein gene. The influence that such genomic instability could have on downstream uses of rescued virus may be detrimental, and we therefore sought to improve the rescue system. Here we report an improved EBOV rescue system with higher efficiency and genome stability, using a modified full-length EBOV clone in Huh7 cells. Moreover, by evaluating a variety of cells lines, we revealed that EBOV genome instability is cell-type dependent, a fact that has significant implications for the preparation of standard virus stocks. Thus, our improved rescue system will have an impact on both basic and translational research in the filovirus field.
KW - Ebola virus
KW - RNA editing site
KW - mutation
KW - reverse genetics system
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U2 - 10.1093/infdis/jiu681
DO - 10.1093/infdis/jiu681
M3 - Article
C2 - 25810440
AN - SCOPUS:84932616092
SN - 0022-1899
VL - 212
SP - S129-S137
JO - Journal of Infectious Diseases
JF - Journal of Infectious Diseases
ER -