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FlyORF-TaDa allows rapid generation of new lines for in vivo cell-type-specific profiling of protein–DNA interactions in Drosophila melanogaster
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ORCID: 0000-0002-3367-7427, McMullen, JPD, Southall, TD and Marshall, OJ ORCID: 0000-0003-1605-3871 2021
, 'FlyORF-TaDa allows rapid generation of new lines for in vivo cell-type-specific profiling of protein–DNA interactions in Drosophila melanogaster'
, G3: Genes, Genomes, Genetics, vol. 11, no. 1
, pp. 1-6
, doi: 10.1093/g3journal/jkaa005.
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(Published version)
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Abstract
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-consumingand 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 color. Profiling the binding of the H3K36me3-associated chromatin protein MRG15 in larval neural stem cells using both FlyORF-TaDa and conventional TaDa demonstrates that newlines generated using this system provide accurate and highly reproducible DamID-binding profiles. Our data further show that MRG15binds 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.
| Item Type: | Article |
|---|---|
| Authors/Creators: | Aughey, GN and Delandre, C and McMullen, JPD and Southall, TD and Marshall, OJ |
| Keywords: | transcription factor, chromatin, transcription, development, DamID, neural stem cells |
| Journal or Publication Title: | G3: Genes, Genomes, Genetics |
| Publisher: | Oxford University Press |
| ISSN: | 2160-1836 |
| DOI / ID Number: | 10.1093/g3journal/jkaa005 |
| Copyright Information: | VC The Author(s) 2020. Published by Oxford University Press on behalf of Genetics Society of America. This is an Open Access article distributed under the terms of the Creative Commons Attribution (CC BY 4.0) License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited |
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