Chromatin and transcription factor profiling in rare stem cell populations using CUT&Tag
| Autor: | Hina Bandukwala, Marjorie Brand, Philippe Gingras-Gelinas, Thomas Olender, Alexandre Blais, Daniel C.L. Robinson, Carmen G. Palii, Yuefeng Li, Kiran Nakka, Odile Neyret, Matthew Man-Kin Wong, F. Jeffrey Dilworth |
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| Rok vydání: | 2021 |
| Předmět: |
flow cytometry/mass cytometry
Science (General) sequence analysis CD34 Mice Transgenic Hindlimb Biology Cleavage (embryo) Cardiotoxins Polymerase Chain Reaction General Biochemistry Genetics and Molecular Biology Histones Mice cell isolation Q1-390 03 medical and health sciences 0302 clinical medicine cell biology Protocol molecular biology Animals Humans Progenitor cell Muscle Skeletal Transcription factor 030304 developmental biology 0303 health sciences General Immunology and Microbiology Stem Cells General Neuroscience sequencing Hematopoietic Stem Cells Chromatin 3. Good health Cell biology Haematopoiesis chromatin immunoprecipitation (ChIP) Stem cell ChIPseq 030217 neurology & neurosurgery Transcription Factors |
| Zdroj: | STAR Protocols, Vol 2, Iss 3, Pp 100751-(2021) STAR Protocols |
| ISSN: | 2666-1667 |
| DOI: | 10.1016/j.xpro.2021.100751 |
| Popis: | Summary Muscle stem cells (MuSCs) are a rare stem cell population that provides myofibers with a remarkable capacity to regenerate after tissue injury. Here, we have adapted the Cleavage Under Target and Tagmentation technology to the mapping of the chromatin landscape and transcription factor binding in 50,000 activated MuSCs isolated from injured mouse hindlimb muscles. We have applied this same approach to human CD34+ hematopoietic stem and progenitor cells. This protocol could be adapted to any rare stem cell population. For complete details on the use and execution of this protocol, please refer to Robinson et al. (2021). Graphical abstract Highlights • Isolation of muscle stem cells from cardiotoxin-injured tissue • CUT&Tag-based mapping of epigenetic landscape and transcription factor enrichment • Adaptation of a synthetic spike-in DNA for effective normalization across samples • Pipeline for efficient analysis of CUT&Tag sequencing data hosted on Git-hub Muscle stem cells (MuSCs) are a rare stem cell population that provides myofibers with a remarkable capacity to regenerate after tissue injury. Here, we have adapted the Cleavage Under Target and Tagmentation technology to the mapping of the chromatin landscape and transcription factor binding in 50,000 activated MuSCs isolated from injured mouse hindlimb muscles. We have applied this same approach to human CD34+ hematopoietic stem and progenitor cells. This protocol could be adapted to any rare stem cell population. |
| Databáze: | OpenAIRE |
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