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![]() 1 publications mentioning mmu-mir-6715Open access articles that are associated with the species Mus musculus and mention the gene name mir-6715. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary. |
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Other miRNAs from this paper: hsa-mir-21, hsa-mir-26a-1, hsa-mir-96, mmu-mir-181a-2, mmu-mir-182, mmu-mir-204, hsa-mir-181a-2, hsa-mir-182, hsa-mir-204, hsa-mir-181a-1, mmu-mir-21a, mmu-mir-26a-1, mmu-mir-96, mmu-mir-181a-1, mmu-mir-26a-2, hsa-mir-26a-2, mmu-mir-21b, mmu-mir-21c, hsa-mir-6715a, hsa-mir-6715b
Using TargetScan, potential targets of miR- 6715-3p and miR-6715-5p were identified, based on the complimentarily of the seed region and 6-7 nt sequence of the target mRNA.
[score:7]
miR-6715-3p and miR-6715-5p were each predicted to target 201 and 206 targets, respectively, with ten overlapping targets (Additional file 5: Table S5).
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To test whether these potential targets have a direct interaction with miR-6715-3p and miR-6715-5p, we examined luciferase activity by inserting the 3’UTR of the potential targets downstream of a luciferase gene.
[score:6]
A suggested pathway for miR-6715-3p and Arhgap12, whereby miR-6715-3p down-regulates expression of Arhgap12, which in turn blocks the hydrolysis of Rac1 (Ras-Related C3 Botulinum Toxin Substrate 1) GTPase.
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The miR-6715 miRNA expression vector was created by cloning the genomic polycistronic miR-6715-3p and miR-6715-5p into the miRvec expression vector (obtained as a gift from Reuven Agami) [43].
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These included protocadherin 19 (Pcdh19) [24] as a candidate target for miR-6715-5p, and vezatin (Vezt) [15], Scn8a [25] and Arhgap12 [16] as candidate targets for miR-6715-3p.
[score:5]
In order to determine what pathway might be impacted by miR-6715-3p and miR-6715-5p regulation, we searched for potential targets.
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Vezt and Arhgap12 were found to be direct targets of miR-6715-3p, with a reduction of luciferase activity by 55% and 15%, respectively (Figure 4B, C).
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Gene targets were identified for each of these miRNAs, including Arhgap12, a GTPase activating protein, for miR-6715-3p, implicating this miRNA in sensory hair cell bundle development, actin reorganization, cell adhesion and inner ear morphogenesis.
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D. Scn8a is not a direct target for miR-6715-3p, as the luciferase activity was not reduced.
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The miR-6715-3p–Arhgap12 miRNA target pair defines a new regulatory pathway.
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miR-6715-3p and miR-6715-5p exhibited increased expression with age between E16 and P8 (*) P < 0.05, (**) P < 0.005. n = 5. B. In situ hybridization of P0 mouse cochlear and vestibular sections.
[score:3]
Protocadherin 19, a member of the δ2 subclass of nonclustered protocadherins and associated with epilepsy and mental retardation [17], is a gene target of miR-6715-5p.
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A. Protocadherin19 as a potential target of miR-6715-5p.
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Figure 2 The genomic location, predicted structure and relative level of expression of miR-6715-3p and miR-6715-5p.
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Vezatin, an adherens junctions transmembrane protein previously implicated in deafness [15] and Arhgap12, a Rho GTPase activating protein [16], were verified to be gene targets of miR-6715-3p.
[score:3]
We validated a number of targets for miR-6715-3p and miR-6715-5p.
[score:3]
Here we suggest a key role for miR-6715-3p in targeting Arhagap12 and in this fashion, influencing Rac1 in the inner ear (Figure 8).
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Figure 3 miR-6715-3p and miR-6715-5p expression in the inner ear.
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B. Vezatin as a potential target of miR-6715-3p.
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In our mo del of the miR-6715-3p circuit in the inner ear, we propose that the miRNA indirectly enhances the action of Rac1 and therefore may promote hair bundle development, actin reorganization, cell adhesion and inner ear morphogenesis.
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miR-6715-3p and miR-6715-5p expression was found in the hair and supporting cells of both cochlea and vestibule, spiral and vestibular ganglia, stria vascularis, basilar membrane and in Reissner's membrane.
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Transfections of these constructs, together with a pre miR-6715 expressing vector into HEK293T cells, and measurement of the relative luciferase activity compared to a mutant 3’UTR construct, were indicative of targeting.
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A. pre-miR6715 is located in the second intron of the Tectb gene.
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Provisional ID chr19_45771, available on miRBase (miR-6715; MI0025026), was detected by an extremely low number of reads (0.0021-0.0126 per million library reads) in oocytes, embryonic day (E)7.5 embryo, E15.5 brain, hippocampus, heart, skin, muscle cells, lung and liver by RNA-seq.
[score:1]
B. miRDeep2 prediction of the RNA secondary hairpin structure of the unprocessed miR-6715-3p, miR-6715-5p, and loop.
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miR-6715-3p and miR-6715-5p were defined for the first time in the inner ear.
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This pri-miRNA produces two mature miRNAs, miR-6715-3p and miR-6715-5p (Figure 2B).
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The seed region of miR-6715-3p is indicated in blue and of miR-6715-5p in green.
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We further validated and characterized the expression of two miRNAs, miR-6715-3p and miR-6715-5p, in the mouse inner ear.
[score:1]
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