Abstract: This present invention relates to methods for improved assays to quantify the level of chromatin targets from biological samples. These assays can be used for the detection of global levels of epigenetic modifications (e.g., histone and DNA modifications), chromatin associated proteins, or chromatin associated ribonucleic acids (RNA). The invention further relates to assay kits that include the reagents needed to prepare biological samples and perform said improved chromatin assays.

Abstract: The invention relates to clinical applications of quantitative chromatin mapping assays, such as chromatin immunoprecipitation assays and assays using tethered enzymes (e.g., chromatin immunocleavage (ChIC) and cleavage under targets & release using nuclease (CUT&RUN®)). The methods may be used to detect and quantitate the presence of epigenetic modifications and mutations on nucleosomes (histones and/or DNA) from biological samples, monitor changes in the status of modifications and mutations, monitor the effectiveness of epigenetic and mutation therapies, select suitable treatments for a disease, determine the prognosis of a subject, identify biomarkers of a disease, and screen for agents that modify epigenetic or mutation status. The invention further relates to kits for use in the methods of the invention.

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.

Abstract: The invention relates to assays for nucleosome remodeling activity using functionalized recombinant mononucleosomes. The functionalized recombinant mononucleosomes comprise a histone octamer comprising recombinant histone H2A, H2B, H3 and H4 proteins and a DNA template comprising a nucleosome positioning sequence that effectively positions the histone octamer and a signal site. The invention further relates to methods of using the assay to quantify enzymatic activity of remodeling enzymes and identifying modulators of remodeling enzyme activity.

Abstract: This present invention relates to methods for carrying out chromatin mapping assays that use enzymes to incorporate barcoded DNA at targeted genomic regions followed by long-read sequencing (e.g., Third Generation Sequencing (TGS)). This approach enables the mapping of chromatin targets using TGS and can be used for a wide range of elements or features, including histone post-translational modifications, chromatin associated proteins, nucleosome positioning, and chromatin accessibility. The invention further relates to kits and reagents for carrying out the methods on chromatin samples that include one or more cells.

Abstract: The present invention relates to DNA-barcoded recombinant nucleosomes and polynucleosomes that have been engineered for use as spike-in controls for the quantitative mapping of chromatin associated proteins using Chromatin ImmunoPrecipitation (ChIP) assays, tethered enzyme-based mapping assays, and other chromatin mapping assays. The invention further relates to methods of using the engineered DNA-barcoded recombinant nucleosomes in ChIP assays, tethered enzyme-based mapping assays, and other chromatin mapping assays.

Abstract: The present invention relates to DNA-barcoded recombinant nucleosomes and polynucleosomes that have been engineered for use as spike-in controls for chromatin accessibility assays, chromatin mapping assays, e.g., using tethered enzymes, as well as other chromatin assays. The invention further relates to methods of using the engineered DNA-barcoded recombinant nucleosomes in chromatin accessibility assays, chromatin mapping assays, as well as other chromatin assays.

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.

Abstract: The invention relates to clinical applications of quantitative chromatin mapping assays, such as chromatin immunoprecipitation assays and assays using tethered enzymes (e.g., chromatin immunocleavage (ChIC) and cleavage under targets & release using nuclease (CUT&RUN®)). The methods may be used to detect and quantitate the presence of epigenetic modifications and mutations on nucleosomes (histones and/or DNA) from biological samples, monitor changes in the status of modifications and mutations, monitor the effectiveness of epigenetic and mutation therapies, select suitable treatments for a disease, determine the prognosis of a subject, identify biomarkers of a disease, and screen for agents that modify epigenetic or mutation status. The invention further relates to kits for use in the methods of the invention.

Abstract: The invention relates to assays for nucleosome remodeling activity using functionalized recombinant mononucleosomes. The functionalized recombinant mononucleosomes comprise a histone octamer comprising recombinant histone H2A, H2B, H3 and H4 proteins and a DNA template comprising a nucleosome positioning sequence that effectively positions the histone octamer and a signal site. The invention further relates to methods of using the assay to quantify enzymatic activity of remodeling enzymes and identifying modulators of remodeling enzyme activity.