Multiplexed single-cell profiling of chromatin states at genomic loci by expansion microscopy
| العنوان: | Multiplexed single-cell profiling of chromatin states at genomic loci by expansion microscopy |
|---|---|
| المؤلفون: | Hao Yuan Kueh, Phuc H.B. Nguyen, Nicholas A. Pease, Kenneth K.H. Ng, Marcus A. Woodworth, Aaron R. Halpern, Joshua C. Vaughan |
| المصدر: | Nucleic Acids Research |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Myosin Light Chains, AcademicSubjects/SCI00010, Cell, RNA polymerase II, Computational biology, Genome, Cell Line, Epigenesis, Genetic, Histones, chemistry.chemical_compound, Narese/3, Genetics, medicine, Humans, Epigenetics, Gene, In Situ Hybridization, Fluorescence, Microscopy, Confocal, biology, Genome, Human, Chromatin, Housekeeping gene, Histone Code, Multicellular organism, medicine.anatomical_structure, Histone, chemistry, Microscopy, Fluorescence, biology.protein, Methods Online, Single-Cell Analysis, DNA |
| الوصف: | Proper regulation of genome architecture and activity is essential for the development and function of multicellular organisms. Histone modifications, acting in combination, specify these activity states at individual genomic loci. However, the methods used to study these modifications often require either a large number of cells or are limited to targeting one histone mark at a time. Here, we developed a new method called Single Cell Evaluation of Post-TRanslational Epigenetic Encoding (SCEPTRE) that uses Expansion Microscopy (ExM) to visualize and quantify multiple histone modifications at non-repetitive genomic regions in single cells at a spatial resolution of ∼75 nm. Using SCEPTRE, we distinguished multiple histone modifications at a single housekeeping gene, quantified histone modification levels at multiple developmentally-regulated genes in individual cells, and evaluated the relationship between histone modifications and RNA polymerase II loading at individual loci. We find extensive variability in epigenetic states between individual gene loci hidden from current population-averaged measurements. These findings establish SCEPTRE as a new technique for multiplexed detection of combinatorial chromatin states at single genomic loci in single cells. Graphical Abstract Graphical AbstractSingle cell evaluation of post-translational epigenetic encoding (SCEPTRE) uses expansion microscopy to combine DNA in situ labeling with immunofluorescence and quantify histone mark levels at individual loci within cells. |
| تدمد: | 1362-4962 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bcf71e7b89e01e9da6c34f28a27bd54f https://pubmed.ncbi.nlm.nih.gov/34048564 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....bcf71e7b89e01e9da6c34f28a27bd54f |
| قاعدة البيانات: | OpenAIRE |
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