citation_mjid: pnas;1913616116v1 og-title: Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system article:published_time: 2019-11-29 twitter:card: summary citation_reference: citation_journal_title=Photochem. Photobiol.;citation_author=M. J. Cormier;citation_author=S. Kuwabara;citation_title=Some observations on the properties of crystalline bacterial luciferase;citation_pages=1217-1225;citation_volume=4;citation_year=1965 citation_journal_title: Proceedings of the National Academy of Sciences type: article citation_author_email: carola.gregor@mpibpc.mpg.de citation_issn: 0027-8424 citation_full_html_url: https://www.pnas.org/content/early/2019/11/27/1913616116.full citation_public_url: https://www.pnas.org/content/early/2019/11/27/1913616116 dc:title: Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system | PNAS Content-Encoding: UTF-8 citation_pdf_url: https://www.pnas.org/content/pnas/early/2019/11/27/1913616116.full.pdf citation_section: Biological Sciences citation_num_pages: 6 citation_fulltext_world_readable: citation_journal_abbrev: PNAS DC.Identifier: 10.1073/pnas.1913616116 DC.Rights: Copyright © 2019 the Author(s). Published by PNAS.. https://creativecommons.org/licenses/by-nc-nd/4.0/This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND). citation_author: Carola Gregor citation_abstract_html_url: https://www.pnas.org/content/early/2019/11/27/1913616116.abstract og-image: HW.identifier: /pnas/early/2019/11/27/1913616116.atom citation_doi: 10.1073/pnas.1913616116 Content-Language: en format-detection: telephone=no Generator: Drupal 7 (http://drupal.org) citation_author_orcid: http://orcid.org/0000-0002-1083-9892 DC.AccessRights: open-access citation_publication_date: 2019/11/29 citation_title: Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system citation_author_institution: Max Planck Institute for Biophysical Chemistry citation_publisher: National Academy of Sciences citation_id: 1913616116v1 og-type: article title: Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system | PNAS og-url: https://www.pnas.org/content/early/2019/11/27/1913616116 DC.Description: Bioluminescence is generated by luciferases that oxidize a specific luciferin. The enzymes involved in the synthesis of the luciferin from widespread cellular metabolites have so far been identified for only 2 bioluminescence systems, those of bacteria and fungi. In these cases, the complete reaction cascade is genetically encodable, meaning that heterologous expression of the corresponding genes can potentially produce autonomous bioluminescence in cell types other than the bacterial or fungal host cells. However, the light levels achieved in mammalian cells so far are not sufficient for single-cell applications. Here we present autonomous bioluminescence images of single mammalian cells by coexpression of the genes encoding the 6 enzymes from the bacterial bioluminescence system. Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterial luxCDABE and frp genes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells. og-description: Bioluminescence is generated by luciferases that oxidize a specific luciferin. The enzymes involved in the synthesis of the luciferin from widespread cellular metabolites have so far been identified for only 2 bioluminescence systems, those of bacteria and fungi. In these cases, the complete reaction cascade is genetically encodable, meaning that heterologous expression of the corresponding genes can potentially produce autonomous bioluminescence in cell types other than the bacterial or fungal host cells. However, the light levels achieved in mammalian cells so far are not sufficient for single-cell applications. Here we present autonomous bioluminescence images of single mammalian cells by coexpression of the genes encoding the 6 enzymes from the bacterial bioluminescence system. Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterial luxCDABE and frp genes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells. Content-Type-Hint: text/html; charset=utf-8 DC.Format: text/html DC.Publisher: National Academy of Sciences DC.Contributor: Carola Gregor Content-Type: application/xhtml+xml; charset=UTF-8 X-Parsed-By: org.apache.tika.parser.DefaultParser article:section: Biological Sciences citation_pmid: 31792180 twitter:title: Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system citation_article_type: Research Article citation_abstract:

Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterial luxCDABE and frp genes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells.

DC.Title: Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system viewport: initial-scale=1, maximum-scale=1, width=device-width, user-scalable=yes HW.pisa: pnas;1913616116v1 DC.Language: en twitter:description: Bioluminescence is generated by luciferases that oxidize a specific luciferin. The enzymes involved in the synthesis of the luciferin from widespread cellular metabolites have so far been identified for only 2 bioluminescence systems, those of bacteria and fungi. In these cases, the complete reaction cascade is genetically encodable, meaning that heterologous expression of the corresponding genes can potentially produce autonomous bioluminescence in cell types other than the bacterial or fungal host cells. However, the light levels achieved in mammalian cells so far are not sufficient for single-cell applications. Here we present autonomous bioluminescence images of single mammalian cells by coexpression of the genes encoding the 6 enzymes from the bacterial bioluminescence system. Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterial luxCDABE and frp genes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells. DC.Date: 2019-12-02 citation_access: all og-site-name: PNAS category: research-article