Peptide ligase-mediated engineering of recombinant nucleosomes
AssigneeEpiCypher, Inc.
InventorsMartis William Cowles, Michael-Christopher Keogh, Jonathan M. Burg, Zu-Wen Sun
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Abstract
The invention relates generally to methods for preparing recombinant nucleosomes. In particular, the invention relates to methods for ligating a histone peptide onto a fully assembled recombinant nucleosome. The invention further relates to modified core histone proteins, histone peptides to be ligated to the modified core histone proteins, and fully assembled recombinant nucleosomes and libraries of recombinant nucleosomes prepared by the methods of the invention.
Claims16
A method for ligating a histone peptide onto a fully assembled recombinant nucleosome, the nucleosome comprising core histones and a polynucleotide wrapped around the core histones;
- the method comprising:
- (a) contacting a histone peptide with a transpeptidase to produce a histone peptide-transpeptidase intermediate, wherein the histone peptide comprises a C-terminal transpeptidase recognition site;
- and
- (b) contacting the histone peptide-transpeptidase intermediate with a fully assembled recombinant nucleosome comprising a modified core histone protein in which the N-terminal tail has been deleted and an amino acid sequence suitable for transpeptidase mediated ligation is added to the N-terminus under conditions sufficient for ligation of the histone peptide to the modified core histone protein;
- thereby producing a fully assembled recombinant nucleosome with a histone peptide ligated thereto.
Dependencies are inferred from the claim text.