FlyORF-TaDa allows rapid generation of new lines for in vivo cell-type specific profiling of protein-DNA interactions in Drosophila melanogaster
Author(s)
Aughey, Gabriel N
Delandre, Caroline
Southall, Tony D
Marshall, Owen J
Type
Working Paper
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-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.
Date Issued
2020-08-06
Citation
2020
Publisher
bioRxiv
Copyright Statement
© 2020 The Author(s). The copyright holder for this preprint (which was not certified by peer review) is the author/funder. It is made available under a CC-BY-NC 4.0 International license http://creativecommons.org/licenses/by-nc/4.0/.
License URL
Sponsor
Wellcome Trust
Biotechnology and Biological Sciences Research Council (BBSRC)
Identifier
https://www.biorxiv.org/content/10.1101/2020.08.06.239251v1
Grant Number
104567/Z/14/Z
BB/P017924/1
Publication Status
Published