FlyORF-TaDa allows rapid generation of new lines for in vivo cell-type specific profiling of protein-DNA interactions in Drosophila melanogaster
File(s)jkaa005.pdf (753.69 KB)
Published version
Author(s)
Southall, Tony
Marshall, Owen J
Aughey, Gabriel N
Delandre, Caroline
McMullen, John PD
Type
Journal Article
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 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 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 26 genome.
Date Issued
2021-01
Date Acceptance
2020-11-07
Citation
G3: Genes, Genomes, Genetics, 2021, 11 (1), pp.1-6
ISSN
2160-1836
Publisher
Genetics Society of America
Start Page
1
End Page
6
Journal / Book Title
G3: Genes, Genomes, Genetics
Volume
11
Issue
1
Copyright Statement
© 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 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.
This is an Open Access article distributed under the terms of the Creative Commons Attribution 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.
License URL
Sponsor
Wellcome Trust
Biotechnology and Biological Sciences Research Council (BBSRC)
Identifier
https://academic.oup.com/g3journal/article/11/1/jkaa005/6044134
Grant Number
104567/Z/14/Z
BB/P017924/1
Subjects
Chromatin
DamID
Development
Neural stem cells
Transcription
Transcription factor
0604 Genetics
Publication Status
Published
Date Publish Online
2020-12-22