Guidelines for minimal reporting requirements, design and interpretation of experiments involving the use of eukaryotic dual gene expression reporters (MINDR)

Abstract

Dual reporters, which encode two distinct proteins within the same mRNA, have played a crucial role in identifying and characterizing new instances of unconventional eukaryotic translation mechanisms. These mechanisms include initiation by internal ribosomal entry sites (IRESs), ribosomal frameshifting, stop codon readthrough, and reinitiation. This design allows one reporter's expression to be influenced by the specific mechanism under investigation, while the other reporter serves as an internal control. However, challenges arise when intervening sequences are placed between these two reporters. Such sequences can inadvertently impact the expression or function of either reporter, independently of translation-related changes. These effects may occur because of cryptic elements inducing or affecting transcription initiation, splicing, polyadenylation, and antisense transcription, as well as unexpected effects of the translated test sequences on the stability and activity of the reporters. Unfortunately, these unintended effects may lead to incorrect conclusions being published in the scientific literature. To address this issue and assist the scientific community in accurately interpreting dual reporter experiments, we have developed comprehensive guidelines. These guidelines cover experiment design, interpretation, and the minimal requirements for reporting results. They are designed to aid researchers conducting these experiments, as well as reviewers, editors, and other investigators who seek to evaluate published data.