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| Status |
Public on Feb 05, 2023 |
| Title |
Comparison of STAMP and TRIBE for identifying the targets of RNA-binding proteins in human and Drosophila cells [Drosophila] |
| Organism |
Drosophila melanogaster |
| Experiment type |
Expression profiling by high throughput sequencing
Other
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| Summary |
RNA binding proteins (RBPs) perform a myriad of functions and are implicated in numerous neurological diseases. To identify the targets of RBPs in small numbers of cells, we developed TRIBE, in which the catalytic domain of the RNA editing enzyme ADAR (ADARcd) is fused to a RBP. In STAMP, the ADARcd is replaced by the RNA editing enzyme Apobec (REF). Here we compared the two enzymes fused to the RBP TDP43 in human cells. Although they both identified TDP43 target mRNAs, combining the two methods more successfully identified high confidence targets. We also assayed the two enzymes in Drosophila cells in which RBP-Apobec fusions generated only low numbers of editing sites comparable to the level of control editing. This was true for two different RBPs, Hrp48 and Thor (Drosophila EIF4E-BP), and contrasted with successful RBP-ADARcd fusions. The results indicate that TRIBE is the method of choice in Drosophila.
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| Overall design |
Comparing TRIBE and STAMP in HEK-293 cells and Drosophila S2 cells. 2 biological replicates for each experiment. Smartseq2 sequencing libraries from cell lines used to identify TRIBE and STAMP induced editing sites.
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| Contributor(s) |
Abruzzi KC, Ratner C, Rosbash M |
| Citation missing |
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| |
| Submission date |
Jan 23, 2023 |
| Last update date |
Feb 06, 2023 |
| Contact name |
Katharine Abruzzi |
| E-mail(s) |
[email protected]
|
| Phone |
7817363161
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| Organization name |
HHMI; Brandeis University
|
| Department |
Biology
|
| Lab |
Rosbash
|
| Street address |
415 South St.
|
| City |
Waltham |
| State/province |
MA |
| ZIP/Postal code |
02478 |
| Country |
USA |
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| Platforms (1) |
| GPL19132 |
Illumina NextSeq 500 (Drosophila melanogaster) |
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| Samples (14)
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| This SubSeries is part of SuperSeries: |
| GSE223557 |
Comparison of STAMP and TRIBE for identifying the targets of RNA-binding proteins in human and Drosophila cells |
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| Relations |
| BioProject |
PRJNA926695 |
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