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| Status |
Public on Jan 31, 2021 |
| Title |
FlyORF-TaDa allows rapid generation of new lines for in vivo cell-type specific profiling of protein-DNA interactions in Drosophila melanogaster |
| Organism |
Drosophila melanogaster |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
Targeted DamID (TaDa) is an increasingly popular method of generating cell-type specific DNA binding profiles in vivo. Although sensitive and versatile, TaDa requires the generation of new transgenic fly lines for every protein that is profiled, which is both time-consuming and costly. Here, we describe the FlyORF-TaDa system for converting an existing FlyORF library of inducible open reading frames (ORFs) to TaDa lines via a genetic cross, with recombinant progeny easily identifiable by eye colour. Profiling the binding of the H3K36me3-associated chromatin protein MRG15 in larval neural stem cells using both FlyORF-TaDa and conventional TaDa demonstrates that new lines generated using this system provide accurate and highly-reproducible DamID binding profiles. Our data further show that MRG15 binds to a subset of active chromatin domains in vivo. Courtesy of the large coverage of the FlyORF library, the FlyORF-TaDa system enables the easy creation of TaDa lines for 74% of all transcription factors and chromatin modifying proteins within the Drosophila genome.
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| Overall design |
Targeted DamID. Two biological replicates for wor-GAL4 driving each of TaDaG2-MRG15, TaDaG2-Dam, FlyORF-TaDa-MRG15, FlyORF-TaDa-Dam.
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| Contributor(s) |
Aughey GN, Delandre C, McMullen JP, Southall TD, Marshall OJ |
| Citation(s) |
33561239 |
| |
| Submission date |
Oct 19, 2020 |
| Last update date |
Apr 13, 2021 |
| Contact name |
Owen Marshall |
| E-mail(s) |
[email protected]
|
| Organization name |
Menzies Institute for Medical Research
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| Street address |
17 Liverpool St
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| City |
Hobart |
| State/province |
Tas |
| ZIP/Postal code |
7000 |
| Country |
Australia |
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| Platforms (2) |
| GPL17275 |
Illumina HiSeq 2500 (Drosophila melanogaster) |
| GPL19132 |
Illumina NextSeq 500 (Drosophila melanogaster) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA669879 |
| SRA |
SRP287684 |
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