Introduction of Dicistrovirus IRESs into UAS/SV40-polyA constructs results in premature polyadenylation and strong overexpression of the upstream ORF in Drosophila animals

Peter V Lidsky; Sergey E Dmitriev; Raul Andino

doi:10.1101/2023.10.04.560905

Introduction of Dicistrovirus IRESs into UAS/SV40-polyA constructs results in premature polyadenylation and strong overexpression of the upstream ORF in Drosophila animals

bioRxiv [Preprint]. 2023 Oct 5:2023.10.04.560905. doi: 10.1101/2023.10.04.560905.

Authors

Peter V Lidsky¹, Sergey E Dmitriev², Raul Andino¹

Affiliations

¹ Department of Microbiology and Immunology, University of California San Francisco, San Francisco, 94158, CA.
² Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119234, Russia.

Abstract

To evaluate the properties of insect virus internal ribosomal entry sites (IRESs) for protein expression in Drosophila, we have introduced Cricket Paralysis virus (CrPV) and Drosophila C virus (DCV) IRESs into UAS/SV40-polyA vector. We found that introduction of IRESs induce premature polyadenylation, resulting in both truncation of the mRNA, and an increase in mRNA levels of approximately 40-fold. The increase in mRNA levels was accompanied by increased resistance to nonsense-mediated mRNA decay (NMD)-mediated degradation. Our results suggest that premature polyadenylation increases mRNA stability in the SV40 polyadenylation site-containing constructs, suggesting a novel method for robust overexpression of transgenes in Drosophila.

Publication types

Preprint

Grants and funding

R01 AI137471/AI/NIAID NIH HHS/United States