Combinatorial single molecule analysis of chromatin
US11479805B2 · kind B2 · utility
Assignees
Inventors
Key dates
| Filing date | Aug 19, 2016 |
| Grant date | Oct 25, 2022 |
| Priority date | — |
| Expiry date | Oct 7, 2038 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N33/6878
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The present invention provides for single-molecule profiling of combinatorial protein modifications and single-molecule profiling of combinatorial protein modifications combined with single-molecule sequencing of protein/nucleic acids complexes. High-throughput single-molecule imaging was applied to decode combinatorial modifications on millions of individual nucleosomes from pluripotent stem cells and lineage-committed cells. Applicants identified bivalent nucleosomes with concomitant repressive and activating marks, as well as other combinatorial modification states whose prevalence varies with developmental potency. Applying genetic and chemical perturbations of chromatin enzymes show a preferential affect on nucleosomes harboring specific modification states. The present invention also combines this proteomic platform with single-molecule DNA sequencing technology to simultaneously determine the modification states and genomic positions of individual nucleosomes. This novel single-molecule technology can be used to address fundamental questions in chromatin biology and epigenetic regulation leading to novel therapeutics and diagnostics.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.