Modular fluorescence complementation sensors for live cell detection of epigenetic signals at endogenous genomic sites

Cristiana Lungu, Sabine Pinter, Julian Broche, Philipp Rathert and Albert Jeltsch ()
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Cristiana Lungu: Stuttgart University
Sabine Pinter: Stuttgart University
Julian Broche: Stuttgart University
Philipp Rathert: Stuttgart University
Albert Jeltsch: Stuttgart University

Nature Communications, 2017, vol. 8, issue 1, 1-13

Abstract: Abstract Investigation of the fundamental role of epigenetic processes requires methods for the locus-specific detection of epigenetic modifications in living cells. Here, we address this urgent demand by developing four modular fluorescence complementation-based epigenetic biosensors for live-cell microscopy applications. These tools combine engineered DNA-binding proteins with domains recognizing defined epigenetic marks, both fused to non-fluorescent fragments of a fluorescent protein. The presence of the epigenetic mark at the target DNA sequence leads to the reconstitution of a functional fluorophore. With this approach, we could for the first time directly detect DNA methylation and histone 3 lysine 9 trimethylation at endogenous genomic sites in live cells and follow dynamic changes in these marks upon drug treatment, induction of epigenetic enzymes and during the cell cycle. We anticipate that this versatile technology will improve our understanding of how specific epigenetic signatures are set, erased and maintained during embryonic development or disease onset.

Date: 2017
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DOI: 10.1038/s41467-017-00457-z

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