Key result
The black queen cell and Drosophila C dicistrovirus intergenic IRES elements were most effective for coupled gene expression in diverse insect cell lines.
The black queen cell and Drosophila C dicistrovirus intergenic IRES elements are highly effective for coupled gene expression in diverse insect cell lines.
May optimize coupled expression in insect systems; leaves open translation to other cell types or applications.
Comparisons of the relative activities of 11 intergenic region (IGR) internal ribosome entry site (IRES) elements of insect dicistrovirus with 5' IRES elements of the hepatitis C and encephalomyocarditis viruses were performed in insect and mammalian cells. Dual luciferase assays were performed to determine the most effective dicistrovirus IGR IRES in the lepidopteran cell lines Sf9 (Spodoptera frugiperda) and BmN (Bombyx mori), and the dipteran cell lines S2 (Drosophila melanogaster) and ATC-10 (Aedes aegypti). Evaluation of dual luciferase expression from DNA plasmids and in vitro-transcribed RNA revealed apparent splicing with certain IRES elements. Though IRES activity depended upon the cell line examined, the black queen cell and Drosophila C dicistrovirus intergenic IRES elements were most effective for coupled gene expression in the diverse insect cell lines examined.
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Carter et al. (2008) studied this question. 11 intergenic region (IGR) internal ribosome entry site (IRES) elements of insect dicistrovirus vs. 5' IRES elements of the hepatitis C and encephalomyocarditis viruses was evaluated on Relative activities (dual luciferase assays). The black queen cell and Drosophila C dicistrovirus intergenic IRES elements were most effective for coupled gene expression in diverse insect cell lines.