Abstract
Genetically encoded FRET-based biosensors are increasingly popular and useful tools for examining signaling pathways with high spatial and temporal resolution in living cells. Here, we show basic techniques used to characterize and to validate single-chain, genetically encoded Förster resonance energy transfer (FRET) biosensors of the Rho GTPase-family proteins. Methods described here are generally applicable to other genetically encoded FRET-based biosensors by modifying the tested conditions to include additional/different regulators and inhibitors, as appropriate for the specific protein of interest.
| Original language | English (US) |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 87-106 |
| Number of pages | 20 |
| DOIs | |
| State | Published - Jan 1 2018 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1821 |
| ISSN (Print) | 1064-3745 |
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Keywords
- Biosensors
- Fluorometry
- FRET
- Ratiometric analysis
- Rho GTPases
- Validation
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Characterization of genetically encoded FRET biosensors for Rho-family GTPases. / Donnelly, Sara K.; Miskolci, Veronika; Garrastegui, Alice M.; Cox, Dianne; Hodgson, Louis.
Methods in Molecular Biology. Humana Press Inc., 2018. p. 87-106 (Methods in Molecular Biology; Vol. 1821).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Characterization of genetically encoded FRET biosensors for Rho-family GTPases
AU - Donnelly, Sara K.
AU - Miskolci, Veronika
AU - Garrastegui, Alice M.
AU - Cox, Dianne
AU - Hodgson, Louis
PY - 2018/1/1
Y1 - 2018/1/1
N2 - Genetically encoded FRET-based biosensors are increasingly popular and useful tools for examining signaling pathways with high spatial and temporal resolution in living cells. Here, we show basic techniques used to characterize and to validate single-chain, genetically encoded Förster resonance energy transfer (FRET) biosensors of the Rho GTPase-family proteins. Methods described here are generally applicable to other genetically encoded FRET-based biosensors by modifying the tested conditions to include additional/different regulators and inhibitors, as appropriate for the specific protein of interest.
AB - Genetically encoded FRET-based biosensors are increasingly popular and useful tools for examining signaling pathways with high spatial and temporal resolution in living cells. Here, we show basic techniques used to characterize and to validate single-chain, genetically encoded Förster resonance energy transfer (FRET) biosensors of the Rho GTPase-family proteins. Methods described here are generally applicable to other genetically encoded FRET-based biosensors by modifying the tested conditions to include additional/different regulators and inhibitors, as appropriate for the specific protein of interest.
KW - Biosensors
KW - Fluorometry
KW - FRET
KW - Ratiometric analysis
KW - Rho GTPases
KW - Validation
UR - http://www.scopus.com/inward/record.url?scp=85051088635&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85051088635&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-8612-5_7
DO - 10.1007/978-1-4939-8612-5_7
M3 - Chapter
C2 - 30062407
AN - SCOPUS:85051088635
T3 - Methods in Molecular Biology
SP - 87
EP - 106
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -