Sustained replication of dengue pseudoinfectious virus lacking the capsid gene by trans-complementation in capsid-producing mosquito cells

• Main Authors: Sangiambut S., Suphatrakul A., Sriburi R., Keelapang P., Puttikhunt C., Kasinrerk W., Malasit P., Sittisombut N.
• Format: Article
• Language: English
• Published: 2014
• Online Access: http://www.scopus.com/inward/record.url?eid=2-s2.0-84877624305&partnerID=40&md5=4e085de9959c293c1a0247634639b8ba; http://www.ncbi.nlm.nih.gov/pubmed/23466247; http://cmuir.cmu.ac.th/handle/6653943832/4049
• Institution: Chiang Mai University

A simple system for the generation of pseudoinfectious particles of dengue virus was developed to facilitate studies of virus replication and vaccine development. Selected clones of the C6/36 mosquito cell line expressing an anchored form of the dengue virus capsid protein served as host cells for the trans-complementation of partially capsid-deleted viral RNA generated in vitro. Transfection of the partially capsid-deleted viral RNA into the anchored capsid-expressing C6/36 cells resulted in moderate titers of infectious virus. Progeny viruses multiplied in the capsid trans-complementing C6/36 cells for up to three weeks, but only initiated single rounds of replication in Vero cells lacking the capsid protein. Employing this trans-complementation system, it was found that nearly all of the capsid-coding sequence in the viral RNA was dispensable for the generation of pseudoinfectious dengue virus particles in mosquito cells. © 2013 Elsevier B.V.