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| Status |
Public on Apr 21, 2021 |
| Title |
Comparison of the specificities of the RGG/RG-domains of FMR1, FXR1 and FXR2 for small RNA |
| Platform organism |
synthetic construct |
| Sample organism |
Homo sapiens |
| Experiment type |
Non-coding RNA profiling by array
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| Summary |
In previous studies, we identified a sequence motif (GDCGG) in almost all microRNAs downregulated in the serum of patients with amyotrophic lateral sclerosis. We found that FMR1, FXR1 and FXR2 directly and specifically interact with microRNAs containing the GDCGG motif via their RGG/RG-domains. Here, we compare the specificities for microRNAs of the RGG/RG-domains of FMR1, FXR1 and FXR2 on a transcriptome-wide scale using the Affymetrix GeneChip™ miRNA 3.0 Arrays.
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| Overall design |
The recombinant RGG/RG-domains of FMR1, FXR1 and FXR2 (2.9 nmole), respectively, were incubated with 50 µg low molecular weight RNA isolated from HEK293 cells in 14 ml of buffer (25 mM Tris, 150 mM potassium chloride, 2.5 mM magnesium chloride, 0.5% NP-40, pH 7.5). After incubation, the recombinant proteins were isolated using Ni-NTA-Agarose (Qiagen #30210), extensively washed and the co-purifying RNA compared to the input-RNA to identify binding microRNAs that get enriched by the different RGG/RG-domains. Three biological replicates were used for each RGG/RG-domain.
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| Contributor(s) |
Freischmidt A, Holzmann K, Weishaupt JH |
| Citation(s) |
33871026 |
| Submission date |
Jul 10, 2017 |
| Last update date |
Jul 25, 2021 |
| Contact name |
Axel Freischmidt |
| E-mail(s) |
axel.freischmidt@uni-ulm.de
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| Phone |
+4973150063051
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| Organization name |
Ulm University
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| Department |
Neurology
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| Street address |
Albert-Einstein-Allee 11
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| City |
Ulm |
| ZIP/Postal code |
89081 |
| Country |
Germany |
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| Platforms (1) |
| GPL16384 |
[miRNA-3] Affymetrix Multispecies miRNA-3 Array |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA393667 |
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