Journal: Genes

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

doi: 10.3390/genes16040444

Figure Lengend Snippet: Figure 1. Development of a dual-color BiAD sensor for detection of histone acetylation. (A) Schematic representation of the sensor design. The target locus is visualized by a full-length fluorophore (YPet), recruited by an MS2 scaffold of the sgRNA in complex with the anchor module (dCas9). One part of the split fluorophore (IFP2.0) is recruited to the SunTag amplification system fused to dCas9 by a single-chain variable fragment (scFv) antibody domain. To detect acetylated histones, a double BRD9 bromodomain (2xBRD9-BD) is fused to the second part of the split IFP2.0 fluorophore. If histone acetylation (Kac) is present at the target locus, the detector module binds at this place bringing the second part of the split IFP2.0 in close spatial proximity to the first one leading to reconstitution of the split IFP2.0 and appearance of fluorescent BiAD signal. (B) Identification of a repetitive genomic region containing histone acetylation, which can be used for the validation of the novel detector module. The gene body of the TTC34 gene contains 392 repeats of the sgRNA binding sites (indicated in the black trace), and it is modified with H3K27ac (indicated in the orange trace).

Article Snippet: The other previously described parts of the dual color BiAD system are available at Addgene (83849).

Techniques: Amplification, Biomarker Discovery, Binding Assay, Modification

Journal: Genes

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

doi: 10.3390/genes16040444

Figure Lengend Snippet: Figure 2. Validation of the BRD9-BD double domain (2xBRD9-BD) for histone acetylation readout at the TTC34 locus. HEK293 cells were transfected with all components of the dual-color BiAD sensor for histone acetylation detection at the TTC34 target locus with either the wildtype 2xBRD9-BD

Article Snippet: The other previously described parts of the dual color BiAD system are available at Addgene (83849).

Techniques: Biomarker Discovery, Transfection

Journal: Genes

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

doi: 10.3390/genes16040444

Figure Lengend Snippet: Figure 3. Visualization of locus-specific gain in histone acetylation upon TSA treatment. HEK293 cells were transfected with all components of the dual-color BiAD sensor for histone acetylation detection

Article Snippet: The other previously described parts of the dual color BiAD system are available at Addgene (83849).

Techniques: Transfection