Single Cell Spatial Chromatin Analysis of Fixed Immunocytochemically Identified Neuronal Cells
| Autor: | James Eberwine, H. Isaac Chen, Jifen Li, Youtao Lu, Jaehee Lee, Jean G. Rosario, Stewart A. Anderson, John A. Wolf, Healy Mimi, Stephen A. Fisher, M. Sean Grady, C. Erik Nordgren, Steven Brem, Alexandra V. Ulyanova, J Wang, Junhyong Kim |
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| Rok vydání: | 2019 |
| Předmět: |
In situ
0303 health sciences Mitochondrial DNA Chemistry Hybridization probe Cell Chromatin Cell biology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Terminator (genetics) medicine.anatomical_structure medicine 030217 neurology & neurosurgery DNA 030304 developmental biology K562 cells |
| Popis: | Assays examining the open-chromatin landscape in single cells require isolation of the nucleus, resulting in the loss of spatial/microenvironment information. Here we describe CHEX-seq (CHromatin EXposed) for identifying single-stranded open-chromatin DNA regions in paraformaldehyde-fixed single cells. CHEX-seq uses light-activated DNA probes that binds to single-stranded DNA in open chromatin. In situ laser activation of the annealed probes’ 3’-Lightning Terminator™ in selected cells permits the probe to act as a primer for in situ enzymatic copying of single-stranded DNA that is then sequenced. CHEX-seq is benchmarked with human K562 cells and its utility is demonstrated in dispersed primary mouse and human brain cells, and immunostained cells in mouse brain sections. Further, CHEX-seq queries the openness of mitochondrial DNA in single cells. Evaluation of an individual cell’s chromatin landscape in its tissue context enables “spatial chromatin analysis”.One Sentence SummaryA new method, CHEX-seq (CHromatin eXposed), identifies the open-chromatin landscape in single fixed cells thereby allowing spatial chromatin analysis of selected cells in complex cellular environments. |
| Databáze: | OpenAIRE |
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