期刊
NATURE METHODS
卷 8, 期 12, 页码 1065-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1763
关键词
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资金
- Swiss National Science Foundation
- SystemsX.ch,
- Marie Curie International Reintegration grant from Seventh Research Framework Programme
- Frontiers in Genetics National Centres of Competence in Research Program
Drosophila melanogaster has one of the best characterized metazoan genomes in terms of functionally annotated regulatory elements. To explore how these elements contribute to gene regulation, we need convenient tools to identify the proteins that bind to them. Here we describe the development and validation of a high-throughput yeast one-hybrid platform, which enables screening of DNA elements versus an array of full-length, sequence-verified clones containing over 85% of predicted Drosophila transcription factors. Using six well-characterized regulatory elements, we identified 33 transcription factor-DNA interactions of which 27 were previously unidentified. To simultaneously validate these interactions and locate the binding sites of involved transcription factors, we implemented a powerful microfluidics-based approach that enabled us to retrieve DNA-occupancy data for each transcription factor throughout the respective target DNA elements. Finally, we biologically validated several interactions and identified two new regulators of sine oculis gene expression and hence eye development.
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