Pré-Publication, Document De Travail Année : 2025

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

1 UCC - University College Cork
2 MSU - Lomonosov Moscow State University = Université d'État Lomonossov de Moscou
3 IBCh RAS - Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
4 Université Paris-Saclay
5 Ǧāmiʿat͏̈ Hayfā = University of Haifa
6 CMU - Carnegie Mellon University [Pittsburgh]
7 CAM - University of Cambridge [Cambridge, UK]
8 UC Davis - University of California [Davis]
9 UHH - Universität Hamburg = University of Hamburg
10 UTokyo - The University of Tokyo
11 UW - Uniwersytet Warszawski [Polska] = University of Warsaw [Poland] = Université de Varsovie [Pologne]
12 Karolinska Institutet = Karolinska Institute [Stockholm]
13 Max Planck Research Group for RNA Biology
14 UIC - University of Illinois [Chicago]
15 HMS - Harvard Medical School [Boston]
16 Hubrecht Institute [Utrecht, Netherlands]
17 TU Delft - Delft University of Technology
18 McGill University = Université McGill [Montréal, Canada]
19 UNITN - Università degli Studi di Trento = University of Trento
20 JMU - Julius-Maximilians-Universität Würzburg = University of Würzburg [Würsburg, Germany]
21 MD Anderson Cancer Center [Houston]
22 UND - University of Notre Dame [Indiana]
23 College of Life Sciences
24 UMD - University of Maryland [College Park]
25 UMB - University of Maryland [Baltimore]
26 Yale University [New Haven]
27 Brigham and Women's Hospital [Boston]
28 University of Colorado Anschutz Medical Campus [Aurora]
29 NYSBC - New York Structural Biology Center
30 UNIBE - Universität Bern = University of Bern = Université de Berne
31 TAMU - Texas A&M University [College Station]
32 Department of Biochemistry
33 Columbia University [New York]
34 University of Alberta
35 MBU / CAS - Institute of Microbiology of the Czech Academy of Sciences [Prague, Czech Republic]
Gary Loughran
  • Fonction : Auteur
  • PersonId : 1335821
Olivier Namy
Pavel V Baranov
  • Fonction : Auteur
  • PersonId : 1335823

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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Dates et versions

hal-04395528 , version 1 (15-01-2024)
hal-04395528 , version 2 (31-01-2025)

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Identifiants

  • HAL Id : hal-04395528 , version 2

Citer

Gary Loughran, Dmitry E Andreev, Ilya M Terenin, Olivier Namy, Martin Mikl, et al.. Guidelines for minimal reporting requirements, design and interpretation of experiments involving the use of eukaryotic dual gene expression reporters (MINDR). 2025. ⟨hal-04395528v2⟩

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