論文 - 詳細
| RRC ID | 64469 |
|---|---|
| 著者 | Buddika K, Xu J, Ariyapala IS, Sokol NS. |
| タイトル | I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells. |
| ジャーナル | Development |
| Abstract |
The adult Drosophila intestinal epithelium is a model system for stem cell biology, but its utility is limited by current biochemical methods that lack cell type resolution. Here, we describe a new proximity-based profiling method that relies upon a GAL4 driver, termed intestinal-kickout-GAL4 (I-KCKT-GAL4), that is exclusively expressed in intestinal progenitor cells. This method uses UV crosslinked whole animal frozen powder as its starting material to immunoprecipitate the RNA cargoes of transgenic epitope-tagged RNA binding proteins driven by I-KCKT-GAL4 When applied to the general mRNA-binder, poly(A)-binding protein, the RNA profile obtained by this method identifies 98.8% of transcripts found after progenitor cell sorting, and has low background noise despite being derived from whole animal lysate. We also mapped the targets of the more selective RNA binder, Fragile X mental retardation protein (FMRP), using enhanced crosslinking and immunoprecipitation (eCLIP), and report for the first time its binding motif in Drosophila cells. This method will therefore enable the RNA profiling of wild-type and mutant intestinal progenitor cells from intact flies exposed to normal and altered environments, as well as the identification of RNA-protein interactions crucial for stem cell function. |
| 巻・号 | 148(1) |
| 公開日 | 2021-1-7 |
| DOI | 10.1242/dev.196568 |
| PII | dev.196568 |
| PMID | 33246929 |
| PMC | PMC7803463 |
| MeSH | Aging / physiology* Animals Base Sequence Drosophila Proteins / metabolism* Drosophila melanogaster / metabolism* Enhancer Elements, Genetic / genetics Female Gene Expression Regulation Genetic Techniques* Intestines / cytology* Organ Specificity Poly(A)-Binding Proteins / metabolism RNA / genetics RNA / metabolism* RNA, Messenger / genetics RNA, Messenger / metabolism Stem Cells / metabolism* Transcription Factors / metabolism* |
| IF | 5.611 |
| 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 | X(Twitter) |
| 各媒体での言及数の合計 | 8 |
| 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 | |
| ショウジョウバエ | DGRC#109126 |