Current Practice in Bicistronic IRES Reporter Use: A Systematic Review.

摘要: Bicistronic reporter assays have been instrumental for transgene expression, understanding of internal ribosomal entry site (IRES) translation, and identification novel cap-independent translational elements (CITE). We observed a large methodological variability in the use bicistronic data presentation or normalization procedures. Therefore, we systematically searched literature IRES studies analyzed details, visualization, Two hundred fifty-seven publications were identified using our search strategy (published 1994-2020). Experimental on eukaryotic adherent cell systems cell-free translation assay included further analysis. evaluated following details 176 full text articles: design, line type, transfection methods, time point analyses post-transfection. For assay, focused methods vitro transcription, type lysate, incubation times temperature. Data can be presented multiple ways: raw from individual cistrons, ratio two, fold changes thereof. In addition, many different control experiments suggested when studying IRES-mediated translation. also categorized summarized their use. Our unbiased provide representative overview parameters that reported inconsistently incompletely, which could hamper reproduction interpretation. On basis analyses, encourage adhering to number practices should improve transparency descriptions facilitate replication.