A General Method to Edit Histone H3 Modifications on Chromatin Via Sortase‐Mediated Metathesis

Abstract The post‐translational modifications (PTMs) on the tail of histone H3 control chromatin structure and influence epigenetics and gene expression. The current chemical methods including unnatural amino acid incorporation and protein splicing enable preparations of the histone with diverse PTMs in cellular contexts, but they are not applicable to edit native chromatin. The manipulation of histone‐modifying enzymes alter the endogenous histone PTMs but the lack of specificity of most histone‐modifying enzymes prevents precise control of specific H3 tail PTM patterns. Here we report a new method to edit the N ‐tail of histone H3 via sortase mediated metathesis (SMM). The sortase can install desired PTM patterns into histone H3 on nucleosomes in vitro and in cellulo . This study expands the application scope of sortase from ligation to metathesis in live cells using cell‐penetrating peptides (CPPs). In addition, it offers a strategy to edit PTMs of cellular histone H3 with potential for the development of precise epigenome editing.