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RHINO comprises a tandem array of three copies of the RNA:DNA hybrid binding domain of human RNase H1 connected by optimized linker segments and fused to a fluorescent protein. This tool allows the measurement of R-loop abundance and dynamics in live cells with high specificity and sensitivity. Using RHINO, we provide a kinetic framework for R-loops at nucleoli, telomeres and protein-coding genes. Our findings demonstrate that R-loop dynamics vary significantly across these regions, potentially reflecting the distinct roles R-loops play in different chromosomal contexts. RHINO is a powerful tool for investigating the role of R-loops in cellular processes and their contribution to disease development and progression.<\/jats:p>","DOI":"10.1093\/nar\/gkad812","type":"journal-article","created":{"date-parts":[[2023,10,11]],"date-time":"2023-10-11T22:35:30Z","timestamp":1697063730000},"page":"11010-11023","source":"Crossref","is-referenced-by-count":16,"title":["Live-cell imaging unveils distinct R-loop populations with heterogeneous dynamics"],"prefix":"10.1093","volume":"51","author":[{"given":"Robert M","family":"Martin","sequence":"first","affiliation":[{"name":"Instituto de Medicina Molecular Jo\u00e3o Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa ,\u00a0 Lisboa , Portugal"}]},{"given":"Madalena R","family":"de\u00a0Almeida","sequence":"additional","affiliation":[{"name":"Instituto de Medicina Molecular Jo\u00e3o Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa ,\u00a0 Lisboa , Portugal"}]},{"given":"Eduardo","family":"Gameiro","sequence":"additional","affiliation":[{"name":"Instituto de Medicina Molecular Jo\u00e3o Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa ,\u00a0 Lisboa , Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7774-1355","authenticated-orcid":false,"given":"S\u00e9rgio F","family":"de\u00a0Almeida","sequence":"additional","affiliation":[{"name":"Instituto de Medicina Molecular Jo\u00e3o Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa ,\u00a0 Lisboa , Portugal"}]}],"member":"286","published-online":{"date-parts":[[2023,10,11]]},"reference":[{"key":"2023111117184497400_B1","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1016\/j.cell.2019.08.055","article-title":"R loops: from physiological to pathological roles","volume":"179","author":"Garc\u00eda-Muse","year":"2019","journal-title":"Cell"},{"key":"2023111117184497400_B2","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1038\/s41580-019-0206-3","article-title":"Regulatory R-loops as facilitators of gene expression and genome stability","volume":"21","author":"Niehrs","year":"2020","journal-title":"Nat. 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