Protocol for identifying genomic binding sites of mitotic bookmarkers in Drosophila neural stem cells and cultured mammalian cells
Summary: Mitotic bookmarkers label key gene loci on highly condensed chromosomes to preserve cell fate memory across mitosis. Here, we present a protocol for identifying binding sites of mitotic bookmarkers (BISMIBs) in developing tissues and cultured cells. We describe a workflow for isolating mito...
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Elsevier
2025-06-01
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725002643 |
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| author | Yuying Shen Jie Liu Yan Song |
| author_facet | Yuying Shen Jie Liu Yan Song |
| author_sort | Yuying Shen |
| collection | DOAJ |
| description | Summary: Mitotic bookmarkers label key gene loci on highly condensed chromosomes to preserve cell fate memory across mitosis. Here, we present a protocol for identifying binding sites of mitotic bookmarkers (BISMIBs) in developing tissues and cultured cells. We describe a workflow for isolating mitotic and interphase cells from Drosophila larval brains and HEK293T cells independent of cell-cycle synchronization. We further detail procedures for low-input in vivo CUT&Tag (Cleavage Under Targets & Tagmentation) sequencing. This protocol is applicable for identifying binding sites for mitotically retained factors in developing tissues.For complete details on the use and execution of this protocol, please refer to Shen et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-7ff420780d1a43fdaf7d9dae0705efe5 |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-7ff420780d1a43fdaf7d9dae0705efe52025-08-20T02:17:28ZengElsevierSTAR Protocols2666-16672025-06-016210385810.1016/j.xpro.2025.103858Protocol for identifying genomic binding sites of mitotic bookmarkers in Drosophila neural stem cells and cultured mammalian cellsYuying Shen0Jie Liu1Yan Song2State Key Laboratory of Membrane Biology, Ministry of Education Key Laboratory of Cell Proliferation and Differentiation, School of Life Sciences, Peking University, Beijing 100871, China; Corresponding authorState Key Laboratory of Membrane Biology, Ministry of Education Key Laboratory of Cell Proliferation and Differentiation, School of Life Sciences, Peking University, Beijing 100871, ChinaState Key Laboratory of Membrane Biology, Ministry of Education Key Laboratory of Cell Proliferation and Differentiation, School of Life Sciences, Peking University, Beijing 100871, China; Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China; Corresponding authorSummary: Mitotic bookmarkers label key gene loci on highly condensed chromosomes to preserve cell fate memory across mitosis. Here, we present a protocol for identifying binding sites of mitotic bookmarkers (BISMIBs) in developing tissues and cultured cells. We describe a workflow for isolating mitotic and interphase cells from Drosophila larval brains and HEK293T cells independent of cell-cycle synchronization. We further detail procedures for low-input in vivo CUT&Tag (Cleavage Under Targets & Tagmentation) sequencing. This protocol is applicable for identifying binding sites for mitotically retained factors in developing tissues.For complete details on the use and execution of this protocol, please refer to Shen et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725002643Sequence analysisCell BiologyCell cultureSingle CellFlow CytometryGenomics |
| spellingShingle | Yuying Shen Jie Liu Yan Song Protocol for identifying genomic binding sites of mitotic bookmarkers in Drosophila neural stem cells and cultured mammalian cells STAR Protocols Sequence analysis Cell Biology Cell culture Single Cell Flow Cytometry Genomics |
| title | Protocol for identifying genomic binding sites of mitotic bookmarkers in Drosophila neural stem cells and cultured mammalian cells |
| title_full | Protocol for identifying genomic binding sites of mitotic bookmarkers in Drosophila neural stem cells and cultured mammalian cells |
| title_fullStr | Protocol for identifying genomic binding sites of mitotic bookmarkers in Drosophila neural stem cells and cultured mammalian cells |
| title_full_unstemmed | Protocol for identifying genomic binding sites of mitotic bookmarkers in Drosophila neural stem cells and cultured mammalian cells |
| title_short | Protocol for identifying genomic binding sites of mitotic bookmarkers in Drosophila neural stem cells and cultured mammalian cells |
| title_sort | protocol for identifying genomic binding sites of mitotic bookmarkers in drosophila neural stem cells and cultured mammalian cells |
| topic | Sequence analysis Cell Biology Cell culture Single Cell Flow Cytometry Genomics |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725002643 |
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