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Selected Works of Miri VanHoven
Article
Bright split red fluorescent proteins for the visualization of endogenous proteins and synapses
Communications Biology (2019)
  • Siyu Feng, University of California - Berkeley
  • Aruna Varshney, San Jose State University
  • Doris Coto Villa, San Jose State University
  • Cyrus Modavi, University of California, San Francisco
  • John Kohler, University of Minnesota
  • Fatima Farah, San Jose State University
  • Shuqin Zhou, University of California, San Francisco
  • Nebat Ali, San Jose State University
  • Joachim D. Müller, University of Minnesota
  • Miri VanHoven, San José State University
  • Bo Huang, University of California, San Francisco
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Abstract
Self-associating split fluorescent proteins (FPs) are split FPs whose two fragments spontaneously associate to form a functional FP. They have been widely used for labeling proteins, scaffolding protein assembly and detecting cell-cell contacts. Recently developments have expanded the palette of self-associating split FPs beyond the original split GFP1-10/11. However, these new ones have suffered from suboptimal fluorescence signal after complementation. Here, by investigating the complementation process, we have demonstrated two approaches to improve split FPs: assistance through SpyTag/SpyCatcher interaction and directed evolution. The latter has yielded two split sfCherry3 variants with substantially enhanced overall brightness, facilitating the tagging of endogenous proteins by gene editing. Based on sfCherry3, we have further developed a new red-colored trans-synaptic marker called Neuroligin-1 sfCherry3 Linker Across Synaptic Partners (NLG-1 CLASP) for multiplexed visualization of neuronal synapses in living C. elegans, demonstrating its broad applications.
Keywords
  • Fluorescence imaging,
  • Fluorescent proteins,
  • Genetic engineering,
  • Neural circuits
Disciplines
Publication Date
September 17, 2019
DOI
10.1038/s42003-019-0589-x
Publisher Statement
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This article was published in Communications Biology, volume 2, issue 1, 2019, and can also be found online here.
© The Author(s) 2019
Citation Information
Siyu Feng, Aruna Varshney, Doris Coto Villa, Cyrus Modavi, et al.. "Bright split red fluorescent proteins for the visualization of endogenous proteins and synapses" Communications Biology Vol. 2 Iss. 1 (2019) ISSN: 23993642
Available at: http://works.bepress.com/miri_vanhoven/14/
Creative Commons license
Creative Commons License
This work is licensed under a Creative Commons CC_BY International License.