High-affinity chromodomains engineered for improved detection of histone methylation and enhanced CRISPR-based gene repression

• DOI: 10.1038/s41467-022-34269-7
• Publication Date: 2022-11-15T15:09:59Z
• Authors: G. Veggiani; R. Villaseñor; G. D. Martyn; J. Q. Tang; M. W. Krone; J. Gu; C. Chen; M. L. Waters; K. H. Pearce; T. Baubec; S. S. Sidhu
• Keywords: 0301 basic medicine; Science; General Biochemistry,Genetics and Molecular Biology; Generation; General Physics and Astronomy; Proteomic analysis; Methylation; Antibodies; Article; Histones; 03 medical and health sciences; Humans; General; Lysine; Q; General Chemistry; Dna; Binding; Chromatin; H3; Domains; Specificity; Phage-display; Nonhistone protein methylation; Protein Processing, Post-Translational

Abstract Histone methylation is an important post-translational modification that plays a crucial role in regulating cellular functions, and its dysregulation implicated cancer developmental defects. Therefore, systematic characterization of histone necessary to elucidate complex biological processes, identify biomarkers, ultimately, enable drug discovery. Studying relies on the use antibodies, but these suffer from lot-to-lot variation, are costly, cannot be used live cells. Chromatin-modification reader domains potential affinity reagents for methylated histones, their application limited by modest affinities. We phage display key residues greatly enhance affinities Cbx chromodomains marks develop general strategy enhancing human protein family. Our allows us powerful probes genome-wide binding analysis live-cell imaging. Furthermore, we optimized extremely potent CRISPR-based repressors tailored gene silencing. results highlight power engineered analyzing interaction networks involving chromatin represent modular platform efficient