Allele-level visualization of transcription and chromatin by high-throughput imaging.
The spatial arrangement of the genome within the nucleus is a pivotal aspect of cellular organization and function with implications for gene expression and regulation. While all genome organization features, such as loops, domains, and radial positioning, are nonrandom, they are characterized by a...
| Publicado en: | Histochemistry & Cell Biology Vol. 162; no. 1/2; pp. 65 - 78 |
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| Autores principales: | , , , |
| Formato: | Journal Article |
| Publicado: |
Springer Nature
Jul2024
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=178293277&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 178293277 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 09486143 NQI jtl: Histochemistry & Cell Biology issn: 09486143 maglogo: N pubinfo: dt: Jul2024 vid: 162 iid: 1/2 pid: 237 pub: Springer Nature place: New York, New York artinfo: ui: 178293277 177123547 10.1007/s00418-024-02289-7 178293277 ppf: 65 ppct: 13 formats: fmt: – @attributes: type: T – @attributes: type: P tig: atl: Allele-level visualization of transcription and chromatin by high-throughput imaging. aug: au: Almansour, Faisal Keikhosravi, Adib Pegoraro, Gianluca Misteli, Tom affil: https://ror.org/040gcmg81 Cell Biology of Genomes, National Cancer Institute, National Institute of Health, 20892, Bethesda, MD, USA sug: ab: The spatial arrangement of the genome within the nucleus is a pivotal aspect of cellular organization and function with implications for gene expression and regulation. While all genome organization features, such as loops, domains, and radial positioning, are nonrandom, they are characterized by a high degree of single-cell variability. Imaging approaches are ideally suited to visualize, measure, and study single-cell heterogeneity in genome organization. Here, we describe two methods for the detection of DNA and RNA of individual gene alleles by fluorescence in situ hybridization (FISH) in a high-throughput format. We have optimized combined DNA/RNA FISH approaches either using simultaneous or sequential detection of DNA and nascent RNA. These optimized DNA and RNA FISH protocols were implemented in a 384-well plate format alongside automated image and data analysis and enable accurate detection of individual gene alleles and their gene expression status across a large cell population. We successfully visualized MYC and EGFR DNA and nascent RNA with allele-level resolution in multiple cell types, and we determined the radial position of active and inactive MYC and EGFR alleles. These optimized DNA/RNA detection approaches are versatile and sensitive tools for mapping of chromatin features and gene activity at the single-allele level and at high throughput. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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