An Improved Reverse Genetics System to Overcome Cell-Type-Dependent Ebola Virus Genome Plasticity.
Yoshimi Tsuda, Thomas Hoenen, Logan Banadyga, Carla Weisend, Stacy M Ricklefs, Stephen F Porcella, Hideki Ebihara
Author Information
Yoshimi Tsuda: Molecular Virology and Host-Pathogen Interaction Unit Department of Microbiology, Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
Thomas Hoenen: Disease Modeling and Transmission Section, Laboratory of Virology.
Logan Banadyga: Molecular Virology and Host-Pathogen Interaction Unit.
Carla Weisend: Molecular Virology and Host-Pathogen Interaction Unit.
Stacy M Ricklefs: Genomics Unit, Research Technology Section, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana.
Stephen F Porcella: Genomics Unit, Research Technology Section, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana.
Hideki Ebihara: Molecular Virology and Host-Pathogen Interaction Unit.
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.
