Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Matryoshka: Ratiometric biosensors from a nested cassette of green- and orange-emitting fluorescent proteins

Cindy Ast, Jessica Foret, Luke M. Oltrogge, Roberto De Michele, Thomas J. Kleist, Cheng-Hsun Ho, Wolf B. Frommer
doi: https://doi.org/10.1101/143107
Cindy Ast
Department of Plant Biology, Carnegie Science, Stanford California 94305, USA;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jessica Foret
Department of Plant Biology, Carnegie Science, Stanford California 94305, USA;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Luke M. Oltrogge
Department of Chemistry, Stanford University, Stanford, California 94305, USA;Current address: Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Roberto De Michele
Institute of Biosciences and Bioresources (IBBR), Italian National Research Council (CNR), Palermo 90129, Italy,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Thomas J. Kleist
Department of Plant Biology, Carnegie Science, Stanford California 94305, USA;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Cheng-Hsun Ho
Department of Plant Biology, Carnegie Science, Stanford California 94305, USA;Agricultural Biotechnology Research Center, Academia Sinica, Academia Road, Nankang, Taipei, Taiwan
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Wolf B. Frommer
Department of Plant Biology, Carnegie Science, Stanford California 94305, USA;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

Sensitivity, dynamic and detection range as well as exclusion of expression and instrumental artifacts are critical for the quantitation of data obtained with fluorescent protein (FP)-based biosensors in vivo. Current biosensors designs are, in general, unable to simultaneously meet all these criteria. Here, we describe a generalizable platform to create dual-FP biosensors with large dynamic ranges by employing a single FP-cassette, named GO-(Green-Orange) Matryoshka. The cassette nests a stable reference FP (large Stokes shift LSSmOrange) within a reporter FP (circularly permuted green FP). GO-Matryoshka yields green and orange fluorescence upon blue excitation. As proof of concept, we converted existing, single-emission biosensors into a series of ratiometric calcium sensors (MatryoshCaMP6s) and ammonium transport activity sensors (AmTryoshka1;3). We additionally identified the internal acid-base equilibrium as a key determinant of the GCaMP dynamic range. Matryoshka technology promises flexibility in the design of a wide spectrum of ratiometric biosensors and expanded in vivo applications.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted June 06, 2017.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Matryoshka: Ratiometric biosensors from a nested cassette of green- and orange-emitting fluorescent proteins
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
Share
Matryoshka: Ratiometric biosensors from a nested cassette of green- and orange-emitting fluorescent proteins
Cindy Ast, Jessica Foret, Luke M. Oltrogge, Roberto De Michele, Thomas J. Kleist, Cheng-Hsun Ho, Wolf B. Frommer
bioRxiv 143107; doi: https://doi.org/10.1101/143107
Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Matryoshka: Ratiometric biosensors from a nested cassette of green- and orange-emitting fluorescent proteins
Cindy Ast, Jessica Foret, Luke M. Oltrogge, Roberto De Michele, Thomas J. Kleist, Cheng-Hsun Ho, Wolf B. Frommer
bioRxiv 143107; doi: https://doi.org/10.1101/143107

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Cell Biology
Subject Areas
All Articles
  • Animal Behavior and Cognition (1529)
  • Biochemistry (2482)
  • Bioengineering (1742)
  • Bioinformatics (9687)
  • Biophysics (3907)
  • Cancer Biology (2974)
  • Cell Biology (4199)
  • Clinical Trials (135)
  • Developmental Biology (2635)
  • Ecology (4104)
  • Epidemiology (2033)
  • Evolutionary Biology (6902)
  • Genetics (5210)
  • Genomics (6508)
  • Immunology (2188)
  • Microbiology (6954)
  • Molecular Biology (2757)
  • Neuroscience (17315)
  • Paleontology (126)
  • Pathology (428)
  • Pharmacology and Toxicology (707)
  • Physiology (1058)
  • Plant Biology (2491)
  • Scientific Communication and Education (645)
  • Synthetic Biology (831)
  • Systems Biology (2690)
  • Zoology (430)