---
_id: '958'
abstract:
- lang: eng
  text: Biosensors that exploit Forster resonance energy transfer (FRET) can be used
    to visualize biological and physiological processes and are capable of providing
    detailed information in both spatial and temporal dimensions. In a FRET-based
    biosensor, substrate binding is associated with a change in the relative positions
    of two fluorophores, leading to a change in FRET efficiency that may be observed
    in the fluorescence spectrum. As a result, their design requires a ligand-binding
    protein that exhibits a conformational change upon binding. However, not all ligand-binding
    proteins produce responsive sensors upon conjugation to fluorescent proteins or
    dyes, and identifying the optimum locations for the fluorophores often involves
    labor-intensive iterative design or high-throughput screening. Combining the genetic
    fusion of a fluorescent protein to the ligand-binding protein with site-specific
    covalent attachment of a fluorescent dye can allow fine control over the positions
    of the two fluorophores, allowing the construction of very sensitive sensors.
    This relies upon the accurate prediction of the locations of the two fluorophores
    in bound and unbound states. In this chapter, we describe a method for computational
    identification of dye-attachment sites that allows the use of cysteine modification
    to attach synthetic dyes that can be paired with a fluorescent protein for the
    purposes of creating FRET sensors.
alternative_title:
- Methods in Molecular Biology
article_processing_charge: No
author:
- first_name: Joshua
  full_name: Mitchell, Joshua
  last_name: Mitchell
- first_name: William
  full_name: Zhang, William
  last_name: Zhang
- first_name: Michel
  full_name: Herde, Michel
  last_name: Herde
- first_name: Christian
  full_name: Henneberger, Christian
  last_name: Henneberger
- first_name: Harald L
  full_name: Janovjak, Harald L
  id: 33BA6C30-F248-11E8-B48F-1D18A9856A87
  last_name: Janovjak
  orcid: 0000-0002-8023-9315
- first_name: Megan
  full_name: O'Mara, Megan
  last_name: O'Mara
- first_name: Colin
  full_name: Jackson, Colin
  last_name: Jackson
citation:
  ama: 'Mitchell J, Zhang W, Herde M, et al. Method for developing optical sensors
    using a synthetic dye fluorescent protein FRET pair and computational modeling
    and assessment. In: Stein V, ed. <i>Synthetic Protein Switches</i>. Vol 1596.
    Synthetic Protein Switches. Springer; 2017:89-99. doi:<a href="https://doi.org/10.1007/978-1-4939-6940-1_6">10.1007/978-1-4939-6940-1_6</a>'
  apa: Mitchell, J., Zhang, W., Herde, M., Henneberger, C., Janovjak, H. L., O’Mara,
    M., &#38; Jackson, C. (2017). Method for developing optical sensors using a synthetic
    dye fluorescent protein FRET pair and computational modeling and assessment. In
    V. Stein (Ed.), <i>Synthetic Protein Switches</i> (Vol. 1596, pp. 89–99). Springer.
    <a href="https://doi.org/10.1007/978-1-4939-6940-1_6">https://doi.org/10.1007/978-1-4939-6940-1_6</a>
  chicago: Mitchell, Joshua, William Zhang, Michel Herde, Christian Henneberger, Harald
    L Janovjak, Megan O’Mara, and Colin Jackson. “Method for Developing Optical Sensors
    Using a Synthetic Dye Fluorescent Protein FRET Pair and Computational Modeling
    and Assessment.” In <i>Synthetic Protein Switches</i>, edited by Viktor Stein,
    1596:89–99. Synthetic Protein Switches. Springer, 2017. <a href="https://doi.org/10.1007/978-1-4939-6940-1_6">https://doi.org/10.1007/978-1-4939-6940-1_6</a>.
  ieee: J. Mitchell <i>et al.</i>, “Method for developing optical sensors using a
    synthetic dye fluorescent protein FRET pair and computational modeling and assessment,”
    in <i>Synthetic Protein Switches</i>, vol. 1596, V. Stein, Ed. Springer, 2017,
    pp. 89–99.
  ista: 'Mitchell J, Zhang W, Herde M, Henneberger C, Janovjak HL, O’Mara M, Jackson
    C. 2017.Method for developing optical sensors using a synthetic dye fluorescent
    protein FRET pair and computational modeling and assessment. In: Synthetic Protein
    Switches. Methods in Molecular Biology, vol. 1596, 89–99.'
  mla: Mitchell, Joshua, et al. “Method for Developing Optical Sensors Using a Synthetic
    Dye Fluorescent Protein FRET Pair and Computational Modeling and Assessment.”
    <i>Synthetic Protein Switches</i>, edited by Viktor Stein, vol. 1596, Springer,
    2017, pp. 89–99, doi:<a href="https://doi.org/10.1007/978-1-4939-6940-1_6">10.1007/978-1-4939-6940-1_6</a>.
  short: J. Mitchell, W. Zhang, M. Herde, C. Henneberger, H.L. Janovjak, M. O’Mara,
    C. Jackson, in:, V. Stein (Ed.), Synthetic Protein Switches, Springer, 2017, pp.
    89–99.
date_created: 2018-12-11T11:49:24Z
date_published: 2017-05-15T00:00:00Z
date_updated: 2025-07-10T12:01:54Z
day: '15'
department:
- _id: HaJa
doi: 10.1007/978-1-4939-6940-1_6
editor:
- first_name: Viktor
  full_name: Stein, Viktor
  last_name: Stein
intvolume: '      1596'
language:
- iso: eng
month: '05'
oa_version: None
page: 89 - 99
publication: Synthetic Protein Switches
publication_identifier:
  issn:
  - 1064-3745
publication_status: published
publisher: Springer
publist_id: '6450'
quality_controlled: '1'
scopus_import: '1'
series_title: Synthetic Protein Switches
status: public
title: Method for developing optical sensors using a synthetic dye fluorescent protein
  FRET pair and computational modeling and assessment
type: book_chapter
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 1596
year: '2017'
...