Development of a BiAD Sensor for Locus-Specific Detection of Cellular Histone Acetylation Dynamics by Fluorescence...

Area of Science:

• Epigenetics
• Molecular Biology
• Cell Biology

Background:

• Dynamic histone acetylation is critical for cellular differentiation and disease.
• Detecting these epigenetic modifications in living cells remains a significant challenge.
• Existing methods lack the ability to track locus-specific acetylation dynamics in real-time.

Purpose of the Study:

• To develop a novel Bimolecular Anchor Detector (BiAD) sensor.
• To enable the detection of locus-specific histone acetylation changes in living cells.
• To utilize fluorescence microscopy for visualizing these dynamic epigenetic events.

Main Methods:

• Engineered a tandem double domain (2xBRD9-BD) bromodomain as a histone acetylation reader.
• Integrated the reader into a dual-color BiAD sensor chassis.
• Identified the TTC34 gene body as a target locus due to dense acetylation and repeats.

Main Results:

• Successfully established a readout of histone acetylation at the TTC34 locus using the BiAD sensor.
• Demonstrated the necessity of the double bromodomain reader for enhanced affinity and specificity.
• Detected dynamic increases in histone acetylation at the TTC34 locus upon Trichostatin A treatment.

Conclusions:

• BiAD sensors utilizing the 2xBRD9-BD reader can effectively detect active chromatin modifications.
• This work expands the utility of BiAD sensors for epigenome studies.
• Highlights the modularity and adaptability of the BiAD sensor platform.