論文 - 詳細
| RRC ID | 46617 |
|---|---|
| 著者 | Jovanovic M, Reiter L, Picotti P, Lange V, Bogan E, Hurschler BA, Blenkiron C, Lehrbach NJ, Ding XC, Weiss M, Schrimpf SP, Miska EA, Grosshans H, Aebersold R, Hengartner MO. |
| タイトル | A quantitative targeted proteomics approach to validate predicted microRNA targets in C. elegans. |
| ジャーナル | Nat Methods |
| Abstract |
Efficient experimental strategies are needed to validate computationally predicted microRNA (miRNA) target genes. Here we present a large-scale targeted proteomics approach to validate predicted miRNA targets in Caenorhabditis elegans. Using selected reaction monitoring (SRM), we quantified 161 proteins of interest in extracts from wild-type and let-7 mutant worms. We demonstrate by independent experimental downstream analyses such as genetic interaction, as well as polysomal profiling and luciferase assays, that validation by targeted proteomics substantially enriched for biologically relevant let-7 interactors. For example, we found that the zinc finger protein ZTF-7 was a bona fide let-7 miRNA target. We also validated predicted miR-58 targets, demonstrating that this approach is adaptable to other miRNAs. We propose that targeted mass spectrometry can be applied generally to validate candidate lists generated by computational methods or in large-scale experiments, and that the described strategy should be readily adaptable to other organisms. |
| 巻・号 | 7(10) |
| ページ | 837-42 |
| 公開日 | 2010-10-1 |
| DOI | 10.1038/nmeth.1504 |
| PII | nmeth.1504 |
| PMID | 20835247 |
| PMC | PMC3444237 |
| MeSH | Animals Base Sequence Caenorhabditis elegans / genetics* Caenorhabditis elegans / metabolism Caenorhabditis elegans Proteins / genetics* Computational Biology / methods Gene Expression Profiling / methods Gene Expression Regulation Genes, Helminth Luciferases / genetics Mass Spectrometry MicroRNAs / genetics* MicroRNAs / metabolism Models, Genetic* Molecular Sequence Data Proteomics / methods* RNA, Messenger / genetics Reverse Transcriptase Polymerase Chain Reaction Species Specificity |
| IF | 30.822 |
| 引用数 | 64 |
| WOS 分野 | BIOCHEMICAL RESEARCH METHODS |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 線虫 | tm600 tm492 |