On the stability of sequences inserted into viral genomes

Willemsen, Anouk; Zwart, Mark P

Only a small number of studies that test the stability of inserts in dsRNA viruses are available, and these concern the generation of recombinant rotavirus expressing foreign genes. Group A rotavirus, consisting of eleven segments, has been engineered to express fluorescent proteins (Kanai et al. 2017 (Kanai et al. , 2019 Komoto et al. 2017 ) such as enhanced green fluorescent protein (eGFP) and mCherry. However, segment 5 in which these genes were introduced is expressed at low levels and is subject to proteasomal degradation ( Based on limited evidence, we tentatively conclude that stoichiometric packaging of segmented genomes may form an impediment to engineering and insert stability. However, recent work also suggests that careful engineering of dsRNA viruses may lead to stable sequence insertions. The generality of these conclusions for the dsRNA viruses, and their dependence on environmental and demographic conditions, remain to be seen.