公开(公告)号：US20230272452A1

公开(公告)日：2023-08-31

申请号：US17989296

申请日：2022-11-17

申请人： The General Hospital Corporation ,  SEQLL INC.

发明人： Efrat Shema-Yaacoby ,  Bradley Bernstein ,  Daniel Jones

IPC分类号： C12Q1/6806 ,  G01N33/68 ,  C12Q1/68

CPC分类号： C12Q1/6806 ,  G01N33/6878 ,  C12Q1/68

摘要： 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.

公开(公告)号：US20240352538A1

公开(公告)日：2024-10-24

申请号：US18764250

申请日：2024-07-04

申请人： Yeda Research and Development Co. Ltd. ,  SeqLL Inc.

发明人： Efrat SHEMA-YAACOBY ,  Erez NIR ,  Vadim FEDYUK ,  Ekaterina ANDREISHCHEVA ,  Daniel JONES ,  Abhijeet SHINDE

IPC分类号： C12Q1/6886 ,  C12Q1/6804

CPC分类号： C12Q1/6886 ,  C12Q1/6804 ,  C12Q2600/156

摘要： A method of analyzing nucleosomes is provided. The method comprising: (a) isolating a plurality of nucleosome molecules from a biological sample; (b) enzymatically linking adenine nucleotides to free DNA ends of the plurality of nucleosome molecules, wherein at least a portion of the adenine nucleotides comprises a label, such that the plurality of nucleosome molecules become attached to a labeled poly(A) tail; (c) hybridizing the plurality of nucleosome molecules attached to the labeled poly(A) tail to a solid support coated with poly(T); and (d) imaging the solid support, whereby the plurality of nucleosome molecules are visualized.