Guidelines for minimal reporting requirements, design and interpretation of experiments involving the use of eukaryotic dual gene expression reporters (MINDR)
Résumé
Dual reporters encoding two distinct proteins within the same mRNA have played a crucial role in identifying and characterizing unconventional mechanisms of eukaryotic translation. These mechanisms include initiation via internal ribosomal entry sites (IRESs), ribosomal frameshifting, stop codon readthrough, and reinitiation. This design enables the expression of one reporter to be influenced by the specific mechanism under investigation, while the other reporter serves as an internal control. However, challenges arise when intervening test sequences are placed between these two reporters. Such sequences can inadvertently impact the expression or function of either reporter, independently of translation-related changes, potentially biasing the results. These effects may occur due to cryptic regulatory elements inducing or affecting transcription initiation, splicing, polyadenylation, and antisense transcription, as well as unpredictable effects of the translated test sequences on the stability and activity of the reporters. Unfortunately, these unintended effects may lead to misinterpretation of the data and the publication of incorrect conclusions 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 experimental 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.
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