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Rational design for superior adenovirus producer cell lines

ITQB and iBET researchers uncover what dictates the productivity of adenovirus vectors cell-lines

Adenovirus vectors (AdV) are efficient gene transfer vectors due to the ability to efficiently infect a wide variety of quiescent and proliferating cell types leading to high-level gene expression. Most replication-defective adenovirus vectors require, for manufacturing and replication, a cell line that expresses the adenoviral E1 functions in trans. E1 region codes for two subunits – E1A and E1B – important to direct cellular and viral gene expression to enable a productive virus cycle. However, the development of these cell lines is typically time demanding in which cells are empirically selected through extensive virus production screening.

During the development of new cell lines for the production of canine adenovirus type 2 vectors, our researchers found that high levels of E1A are proportional to virus amplification, while high E1B expression prolonged cell viability, contributing differently to the global viral titer. E1B expression is even more relevant in the context of helper dependent vector production where the cytotoxicity of Cre recombinase must be counteracted. These findings highlight the ability to express high levels of E1A and E1B as an important feature to increase both the adenovirus yields and quality.

Original Article

PLoS One. 2013;8(4)

Impact of E1 and Cre on Adenovirus Vector Amplification: Developing MDCK CAV-2-E1 and E1-Cre Transcomplementing Cell Lines

Paulo Fernandes, Virgínia M. Santiago, Ana F. Rodrigues, Hélio Tomás, Eric J. Kremer,
Paula M. Alves and Ana S. Coroadinha

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