Evidence map›Paper›PMID 29921790›Full record

ArticleGenes2018

MiRAR-miRNA Activity Reporter for Living Cells.

Matthew A Turk, Christina Z Chung, Emad Manni, Stephanie A Zukowski, Anish Engineer, Yasaman Badakhshi, Yumin Bi, Ilka U Heinemann

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Genes, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.

0numbers the graph read from it
0cells of the map it votes in
8citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

8 citing papers in PubMed.

  1. Article
  2. Article
  3. Review
  4. Review
  5. Article
  6. Review
  7. Article
  8. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors.

Matthew A TurkDepartment of Biochemistry, The University of Western Ontario, 1151 Richmond Street, London, ON N6A 5C1, Canada. mturk5@uwo.ca.
Christina Z ChungDepartment of Biochemistry, The University of Western Ontario, 1151 Richmond Street, London, ON N6A 5C1, Canada. cchung88@uwo.ca.
Emad ManniDepartment of Biochemistry, The University of Western Ontario, 1151 Richmond Street, London, ON N6A 5C1, Canada. emanni@uwo.ca.
Stephanie A ZukowskiDepartment of Biochemistry, The University of Western Ontario, 1151 Richmond Street, London, ON N6A 5C1, Canada. szukowsk@uwo.ca.
Anish EngineerDepartment of Physiology and Pharmacology, The University of Western Ontario, 1151 Richmond Street, London, ON N6A 5C1, Canada. aengine@uwo.ca.
Yasaman BadakhshiDepartment of Biochemistry, The University of Western Ontario, 1151 Richmond Street, London, ON N6A 5C1, Canada. ybadakhs@uwo.ca.
Yumin BiDepartment of Biochemistry, The University of Western Ontario, 1151 Richmond Street, London, ON N6A 5C1, Canada. ybi@uwo.ca.
Ilka U HeinemannDepartment of Biochemistry, The University of Western Ontario, 1151 Richmond Street, London, ON N6A 5C1, Canada. ilka.heinemann@uwo.ca.com.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

microRNA (miRNA) activity and regulation are of increasing interest as new therapeutic targets. Traditional approaches to assess miRNA levels in cells rely on RNA sequencing or quantitative PCR. While useful, these approaches are based on RNA extraction and cannot be applied in real-time to observe miRNA activity with single-cell resolution. We developed a green fluorescence protein (GFP)-based reporter system that allows for a direct, real-time readout of changes in miRNA activity in live cells. The miRNA activity reporter (MiRAR) consists of GFP fused to a 3′ untranslated region containing specific miRNA binding sites, resulting in miRNA activity-dependent GFP expression. Using qPCR, we verified the inverse relationship of GFP fluorescence and miRNA levels. We demonstrated that this novel optogenetic reporter system quantifies cellular levels of the tumor suppressor miRNA let-7 in real-time in single Human embryonic kidney 293 (HEK 293) cells. Our data shows that the MiRAR can be applied to detect changes in miRNA levels upon disruption of miRNA degradation pathways. We further show that the reporter could be adapted to monitor another disease-relevant miRNA, miR-122. With trivial modifications, this approach could be applied across the miRNome for quantification of many specific miRNA in cell cultures, tissues, or transgenic animal models.

Indexed as

fluorescent reporterlive cell imagingmicroRNA quantificationoptogeneticssmall molecule drug screening

Identifiers

PMID29921790
PMCPMC6027049

What OpenQuestion holds

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Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.