Flaviviruses such as the dengue (DENV) and the Zika virus (ZIKV) are important human pathogens causing around 100 million symptomatic infections each year. During infection, small subgenomic flavivirus RNAs (sfRNAs) are formed inside the infected host cell as a result of incomplete degradation of the viral RNA genome by cellular exoribonuclease XRN1. Although the full extent of sfRNA functions is to be revealed, these non-coding RNAs are key virulence factors and their detrimental effects on multiple cellular processes seem to consistently involve molecular interactions with RNA-binding proteins (RBPs). Discovery of such sfRNA-binding host-factors has followed established biochemical pull-down approaches skewed towards highly abundant proteins hampering proteome-wide coverage. Yeast three-hybrid (Y3H) systems represent an attractive alternative approach. To facilitate proteome-wide screens for RBP, we revisited and improved existing RNA-Y3H methodology by (1) implementing full-length ORF libraries in combination with (2) efficient yeast mating to increase screening depth and sensitivity, and (3) stringent negative controls to eliminate over-representation of non-specific RNA-binders. These improvements were validated employing the well-characterized interaction between DDX6 (DEAD-box helicase 6) and sfRNA of DENV as paradigm. Our advanced Y3H system was used to screen for human proteins binding to DENV and ZIKV sfRNA, resulting in a list of 69 putative sfRNA-binders, including several previously reported as well as numerous novel RBP host factors. Our methodology requiring no sophisticated infrastructure or analytic pipeline may be employed for the discovery of meaningful RNA–protein interactions at large scale in other fields.
The recent decline in vaccination rates across Europe has led to outbreaks of vaccine preventable diseases such as measles. Although there have always been persons opposed to vaccination, in recent years an increasing number of parents are losing confidence in or are being complacent about vaccinating their children. Given the accessibility of information on the internet, parents are actively and independently researching vaccines. They are exposed to negative claims about vaccines that appeal to their emotions and emotional stories tend make parents doubt vaccinations. By contrast, most positive vaccine messages focus on providing information through scientific data which has proven to be ineffective for some hesitant parents.Vaccine hesitancy can only be understood by looking at it from different perspectives and by exchanging knowledge between multiple fields of study. A transdisciplinary approach, in which individuals with different backgrounds search for solutions together, is necessary to be able to provide one or more solutions to the problem. Therefore, we spent eight months trying to solve part of this wicked <target target-type="page-num" id="p-93"/>problem from a transdisciplinary perspective. Based on literature reviews of different topics within vaccine hesitancy, interviews with hesitant parents, interactions with various stakeholders within and outside academia, and analyses of popular views on social media concerning vaccines, we question the effectiveness of the current pro-vaccine approaches. We also suggest adopting ‘storytelling’ that incorporates scientific data to inform parents and we argue that narratives are intrinsically persuasive as they are easier to understand and could prove more effective than traditional scientific communication. We expect that this strategy will contribute to the increase in and maintenance of high vaccination coverage rates and stop the circulation and outbreak of vaccine preventable diseases.
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