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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

Citation

Aughey, GN and Delandre, C and McMullen, JPD and Southall, TD and Marshall, OJ, 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, 11, (1) pp. 1-6. ISSN 2160-1836 (2021) [Refereed Article]


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Copyright Statement

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

DOI: doi:10.1093/g3journal/jkaa005

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 genome.

Item Details

Item Type:Refereed Article
Keywords:transcription factor, chromatin, transcription, development, DamID, neural stem cells
Research Division:Biological Sciences
Research Group:Genetics
Research Field:Epigenetics (incl. genome methylation and epigenomics)
Objective Division:Health
Objective Group:Clinical health
Objective Field:Treatment of human diseases and conditions
UTAS Author:Delandre, C (Dr Caroline Delandre)
UTAS Author:McMullen, JPD (Mr John McMullen)
UTAS Author:Marshall, OJ (Dr Owen Marshall)
ID Code:146163
Year Published:2021
Web of Science® Times Cited:1
Deposited By:Menzies Institute for Medical Research
Deposited On:2021-08-24
Last Modified:2022-08-19
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