Direct observation of cytoskeleton-dependent trafficking of miRNA visualized by the introduction of pre-miRNA
Summary: MicroRNA (miRNA) plays physiologically and pathologically important roles in post-transcriptional regulation. Although miRNA has been suggested to dynamically interact with cellular organelles, the dynamicity of intracellular miRNA behavior has remained unclear. Here, by introducing fluores...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-02-01
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| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004224000324 |
| _version_ | 1827377923051487232 |
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| author | Toshinari Ishikawa Ko Sugawara Junwei Zhang Takashi Funatsu Kohki Okabe |
| author_facet | Toshinari Ishikawa Ko Sugawara Junwei Zhang Takashi Funatsu Kohki Okabe |
| author_sort | Toshinari Ishikawa |
| collection | DOAJ |
| description | Summary: MicroRNA (miRNA) plays physiologically and pathologically important roles in post-transcriptional regulation. Although miRNA has been suggested to dynamically interact with cellular organelles, the dynamicity of intracellular miRNA behavior has remained unclear. Here, by introducing fluorescently labeled pre-miRNA into living cells, we improved the miRNA visualization method using exogenous miRNA precursors. Through the combination of our miRNA visualization method and single-molecule sensitive fluorescence microscopy, we quantitatively analyzed the process of miRNA maturation. Furthermore, single-particle tracking of fluorescent miRNA in cells revealed the directed movements of miRNA on cytoskeletal components (i.e., microtubules and actin filaments). Our results also suggest that cytoskeleton-dependent miRNA trafficking is associated with the interaction of miRNAs with the nucleus and the endoplasmic reticulum/Golgi apparatus. Our method should facilitate the elucidation of the mechanism and physiological significance of the subcellular localization and organelle interaction of miRNA. |
| first_indexed | 2024-03-08T12:47:25Z |
| format | Article |
| id | doaj.art-803a9052d11a4a439521df6331986a5b |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-03-08T12:47:25Z |
| publishDate | 2024-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-803a9052d11a4a439521df6331986a5b2024-01-21T05:09:53ZengElsevieriScience2589-00422024-02-01272108811Direct observation of cytoskeleton-dependent trafficking of miRNA visualized by the introduction of pre-miRNAToshinari Ishikawa0Ko Sugawara1Junwei Zhang2Takashi Funatsu3Kohki Okabe4Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanGraduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanGraduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, JapanGraduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan; Corresponding authorGraduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan; JST, PRESTO, 4-8-1 Honcho, Kawaguchi, Saitama 332-0012, Japan; Corresponding authorSummary: MicroRNA (miRNA) plays physiologically and pathologically important roles in post-transcriptional regulation. Although miRNA has been suggested to dynamically interact with cellular organelles, the dynamicity of intracellular miRNA behavior has remained unclear. Here, by introducing fluorescently labeled pre-miRNA into living cells, we improved the miRNA visualization method using exogenous miRNA precursors. Through the combination of our miRNA visualization method and single-molecule sensitive fluorescence microscopy, we quantitatively analyzed the process of miRNA maturation. Furthermore, single-particle tracking of fluorescent miRNA in cells revealed the directed movements of miRNA on cytoskeletal components (i.e., microtubules and actin filaments). Our results also suggest that cytoskeleton-dependent miRNA trafficking is associated with the interaction of miRNAs with the nucleus and the endoplasmic reticulum/Golgi apparatus. Our method should facilitate the elucidation of the mechanism and physiological significance of the subcellular localization and organelle interaction of miRNA.http://www.sciencedirect.com/science/article/pii/S2589004224000324Nucleic acidsMolecular physiologyMolecular biologyBiomechanics |
| spellingShingle | Toshinari Ishikawa Ko Sugawara Junwei Zhang Takashi Funatsu Kohki Okabe Direct observation of cytoskeleton-dependent trafficking of miRNA visualized by the introduction of pre-miRNA iScience Nucleic acids Molecular physiology Molecular biology Biomechanics |
| title | Direct observation of cytoskeleton-dependent trafficking of miRNA visualized by the introduction of pre-miRNA |
| title_full | Direct observation of cytoskeleton-dependent trafficking of miRNA visualized by the introduction of pre-miRNA |
| title_fullStr | Direct observation of cytoskeleton-dependent trafficking of miRNA visualized by the introduction of pre-miRNA |
| title_full_unstemmed | Direct observation of cytoskeleton-dependent trafficking of miRNA visualized by the introduction of pre-miRNA |
| title_short | Direct observation of cytoskeleton-dependent trafficking of miRNA visualized by the introduction of pre-miRNA |
| title_sort | direct observation of cytoskeleton dependent trafficking of mirna visualized by the introduction of pre mirna |
| topic | Nucleic acids Molecular physiology Molecular biology Biomechanics |
| url | http://www.sciencedirect.com/science/article/pii/S2589004224000324 |
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