Sci Rep
 2017
DOI: 10.1038/srep45197
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Dissecting miRNA gene repression on single cell level with an advanced fluorescent reporter system

Nicolás Lemus-Diaz
1
,
Kai O. Böker
2
,
Ignacio Rodríguez-Polo
3
et al.

Abstract: Despite major advances on miRNA profiling and target predictions, functional readouts for endogenous miRNAs are limited and frequently lead to contradicting conclusions. Numerous approaches including functional high-throughput and miRISC complex evaluations suggest that the functional miRNAome differs from the predictions based on quantitative sRNA profiling. To resolve the apparent contradiction of expression versus function, we generated and applied a fluorescence reporter gene assay enabling single cell ana… Show more

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Cited by 18 publications

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“…Consistently, based on a mathematical model‐derived predictions to describe gene regulation on the single cell level, another pioneer study had previously described that low expressed miRNA below median have low or no inhibitory capability, while miRNAs over the median and third quartile show a range of behavior that does not reflect their expression levels (Lemus Diaz et al , 2017 ). Of note, the authors propose that the gene regulatory function is actually a dynamic and complex process, in which some miRNAs also display their functionality according to the subcellular location, with low functional and high expressed miRNA being at least partially located in the nucleus, while some mid expressed but high functional miRNAs are mainly seen in the cytoplasmic region (Lemus Diaz et al , 2017 ).…”
Section: Resultsmentioning
confidence: 92%
Exaggerated anticipatory anxiety is common in social anxiety disorder (SAD). Neuroimaging studies have revealed altered neural activity in response to social stimuli in SAD, but fewer studies have examined neural activity during anticipation of feared social stimuli in SAD. The current study examined the time course and magnitude of activity in threat processing brain regions during speech anticipation in socially anxious individuals and healthy controls (HC). Method Participants (SAD n = 58; HC n = 16) underwent functional magnetic resonance imaging (fMRI) during which they completed a 90s control anticipation task and 90s speech anticipation task.
“…Consistently, based on a mathematical model‐derived predictions to describe gene regulation on the single cell level, another pioneer study had previously described that low expressed miRNA below median have low or no inhibitory capability, while miRNAs over the median and third quartile show a range of behavior that does not reflect their expression levels (Lemus Diaz et al , 2017 ). Of note, the authors propose that the gene regulatory function is actually a dynamic and complex process, in which some miRNAs also display their functionality according to the subcellular location, with low functional and high expressed miRNA being at least partially located in the nucleus, while some mid expressed but high functional miRNAs are mainly seen in the cytoplasmic region (Lemus Diaz et al , 2017 ).…”
Section: Resultsmentioning
confidence: 92%
Exaggerated anticipatory anxiety is common in social anxiety disorder (SAD). Neuroimaging studies have revealed altered neural activity in response to social stimuli in SAD, but fewer studies have examined neural activity during anticipation of feared social stimuli in SAD. The current study examined the time course and magnitude of activity in threat processing brain regions during speech anticipation in socially anxious individuals and healthy controls (HC). Method Participants (SAD n = 58; HC n = 16) underwent functional magnetic resonance imaging (fMRI) during which they completed a 90s control anticipation task and 90s speech anticipation task.
“…Comparison of the relative YFP (control) and CFP (target) expression in the cell population then enables the efficiency of each miRNA to be assessed. It has previously been shown with this system, that low proficiency miRNAs decrease the expression of CFP relative to YFP, while maintaining a linear relationship between expression of the two fluorophores, leading to only a subtle shift in the flow cytometry profile ( 27 ). In contrast, the action of high proficiency miRNAs leads to a non-linear relationship between CFP and YFP expression, and formation of strongly shifted, curved profiles.…”
Section: Resultsmentioning
confidence: 99%
Exaggerated anticipatory anxiety is common in social anxiety disorder (SAD). Neuroimaging studies have revealed altered neural activity in response to social stimuli in SAD, but fewer studies have examined neural activity during anticipation of feared social stimuli in SAD. The current study examined the time course and magnitude of activity in threat processing brain regions during speech anticipation in socially anxious individuals and healthy controls (HC). Method Participants (SAD n = 58; HC n = 16) underwent functional magnetic resonance imaging (fMRI) during which they completed a 90s control anticipation task and 90s speech anticipation task.
“…To verify the biological activity linked to miRNAs enriched in WJ-MSC-derived EV fractions, we used the untranslated trans assay (UTA) dual-fluorescence reporter system, 34 where the cyan fluorescent protein (CFP) gene carries a selected miRNA complementary sequence that, if present, alters its fluorescence level ( Figure 4 A). Briefly, UTA expression plasmids (pUTA) were constructed by inserting complementary regions for miR-146a-5p, miR-423-3p, miR-423-5p, miR-92b-3p, and miR-191-5p within the 3′ UTR of CFP.…”
Section: Resultsmentioning
confidence: 99%
Exaggerated anticipatory anxiety is common in social anxiety disorder (SAD). Neuroimaging studies have revealed altered neural activity in response to social stimuli in SAD, but fewer studies have examined neural activity during anticipation of feared social stimuli in SAD. The current study examined the time course and magnitude of activity in threat processing brain regions during speech anticipation in socially anxious individuals and healthy controls (HC). Method Participants (SAD n = 58; HC n = 16) underwent functional magnetic resonance imaging (fMRI) during which they completed a 90s control anticipation task and 90s speech anticipation task.
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