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Photocaged Hoechst Enables Subnuclear Visualization and Cell Selective Staining of DNA in vivo |
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| Authors: | Carina A Lämmle Adam Varady Thorsten G Müller Dr Caterina Sturtzel Michael Riepl Bettina Mathes Jenny Eichhorst Dr Anje Sporbert Dr Martin Lehmann Prof?Dr Hans-Georg Kräusslich Dr Martin Distel Dr Johannes Broichhagen |
| Affiliation: | 1. Department of Chemical Biology, Max Planck Institute for Medical Research, Jahnstr. 29, 69120 Heidelberg, Germany;2. St. Anna Children's Cancer Research Institute, Innovative Cancer Models, Zimmermannplatz 10, 1090 Vienna, Austria
These authors contributed equally to this work.;3. Department of Infectious Diseases, Virology, University Hospital Heidelberg, Im Neuenheimer Feld 344, 69120 Heidelberg, Germany;4. St. Anna Children's Cancer Research Institute, Innovative Cancer Models, Zimmermannplatz 10, 1090 Vienna, Austria Zebrafish Platform Austria for preclinical drug screening (ZANDR), Zimmermannplatz 10, 1090 Vienna, Austria;5. St. Anna Children's Cancer Research Institute, Innovative Cancer Models, Zimmermannplatz 10, 1090 Vienna, Austria;6. Department of Molecular Pharmacology and Cell Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Straße 10, 13125 Berlin, Germany;7. Advanced Light Microscopy, Max Delbrück Centrum for Molecular Medicine Berlin in the Helmholtz Association, Robert-Rössle-Straße 10, 13125 Berlin, Germany |
| Abstract: | Selective targeting of DNA by means of fluorescent labeling has become a mainstay in the life sciences. While genetic engineering serves as a powerful technique and allows the visualization of nucleic acid by using DNA-targeting fluorescent fusion proteins in a cell-type- and subcellular-specific manner, it relies on the introduction of foreign genes. On the other hand, DNA-binding small fluorescent molecules can be used without genetic engineering, but they are not spatially restricted. Herein, we report a photocaged version of the DNA dye Hoechst33342 (pcHoechst), which can be uncaged by using UV to blue light for the selective staining of chromosomal DNA in subnuclear regions of live cells. Expanding its application to a vertebrate model organism, we demonstrate uncaging in epithelial cells and short-term cell tracking in vivo in zebrafish. We envision pcHoechst as a valuable tool for targeting and interrogating DNA with precise spatiotemporal resolution in living cells and wild-type organisms. |
| Keywords: | DNA Hoechst labeling microscopy zebrafish |