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Updated: May 23, 2025
24:02
The ChroP Approach Combines ChIP and Mass Spectrometry to Dissect Locus-specific Proteomic Landscapes of Chromatin
Published on: April 11, 2014
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Coupling Proximity Biotinylation with Genomic Targeting to Characterize Locus-Specific Changes in Chromatin
Pata-Eting Kougnassoukou Tchara1,2,3, Jérémy Loehr1,2,3, Jean-Philippe Lambert1,2,3
1Department of Molecular Medicine and Cancer Research Centre, Université Laval, Quebec, QC G1V 0A6, Canada.
Journal of Proteome Research
|March 7, 2025
Summary
New CRISPR-based tools, CasTurbo and CasUltra, enable locus-specific proteomics to map chromatin changes. These tools reveal how inhibiting bromodomain and extra-terminal (BET) proteins impacts the MYC promoter in melanoma cells.
Area of Science:
- Genomics
- Proteomics
- Molecular Biology
Background:
- Gene expression regulation relies on chromatin modifications at specific loci.
- Understanding these changes during drug treatment or disease is challenging.
Purpose of the Study:
- To develop novel tools for locus-specific chromatin proteomics.
- To investigate the impact of bromodomain inhibitor JQ1 on the MYC promoter in melanoma.
Main Methods:
- CRISPR/Cas9 genome targeting combined with TurboID/UltraID proximity biotinylation.
- Application of CasUltra for proteomic profiling of the MYC promoter in A375 melanoma cells.
- Quantification of chromatin protein changes upon JQ1 treatment.
Main Results:
- CasTurbo and CasUltra efficiently labeled chromatin proteins at repetitive and non-amplified gene loci.
- JQ1 treatment caused significant reorganization of chromatin composition at the MYC promoter.
- BET protein displacement and retention correlated with JQ1 response.
Conclusions:
- CasTurbo and CasUltra are versatile tools for in vivo, locus-specific proteome profiling.
- These tools provide insights into the molecular mechanisms of gene regulation and drug response.
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