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

Loughran, G.; Andreev, D.E.; Terenin, I.M.; Namy, O.; Mikl, M.; Yordanova, M.M.; McManus, C.J.; Firth, A.E.; Atkins, J.F.; Fraser, C.S.; Ignatova, Z.; Iwasaki, S.; Kufel, J.; Larsson, O.; Leidel, S.A.; Mankin, A.S.; Mariotti, M.; Tanenbaum, M.E.; Topisirovic, I.; Vázquez-Laslop, N.; Viero, G.; Caliskan, N.; Chen, Y.; Clark, P.L.; Dinman, J.D.; Farabaugh, P.J.; Gilbert, W.V.; Ivanov, P.; Kieft, J.S.; Mühlemann, O.; Sachs, M.S.; Shatsky, I.N.; Sonenberg, N.; Steckelberg, A.; Willis, A.E.; Woodside, M.T.; Valasek, L.S.; Dmitriev, S.E.; Baranov, P.V.

Авторы: Loughran Gary, Andreev Dmitry E., Terenin Ilya M., Namy Olivier, Mikl Martin, Yordanova Martina M., McManus C.Joel, Firth Andrew E., Atkins John F., Fraser Christopher S., Ignatova Zoya, Iwasaki Shintaro, Kufel Joanna, Larsson Ola, Leidel Sebastian A., Mankin Alexander S., Mariotti Marco, Tanenbaum Marvin E., Topisirovic Ivan, Vázquez-Laslop Nora, Viero Gabriela, Caliskan Neva, Chen Yiwen, Clark Patricia L., Dinman Jonathan D., Farabaugh Philip J., Gilbert Wendy V., Ivanov Pavel, Kieft Jeffrey S., Mühlemann Oliver, Sachs Matthew S., Shatsky Ivan N., Sonenberg Nahum, Steckelberg Anna-Lena, Willis Anne E., Woodside Michael T., Valasek Leos Shivaya, Dmitriev Sergey E., Baranov Pavel V.
Журнал: Nature Structural and Molecular Biology
Том: 32
Номер: 3
Год издания: 2025
Издательство: Nature Publishing Group
Местоположение издательства: United Kingdom
Первая страница: 418
Последняя страница: 430
DOI: 10.1038/s41594-025-01492-x
Аннотация: Dual reporters encoding two distinct proteins within the same mRNA have had 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, independent 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 data and the publication of incorrect conclusions in the scientific literature. To address this issue and to 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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