Single-Molecule FRET Assay to Observe the Activity of Proteins Involved in RNA/RNA Annealing.
Methods Mol Biol
; 1737: 301-319, 2018.
Article
in En
| MEDLINE
| ID: mdl-29484600
ABSTRACT
In recent years, single-molecule fluorescence resonance energy transfer (smFRET) has emerged as a powerful technique to study macromolecular interactions. The chief advantages of smFRET analysis compared to bulk measurements include the possibility to detect sample heterogeneities within a large population of molecules and the facility to measure kinetics without needing the synchronization of intermediate states. As such, the methodology is particularly well adapted to observe and analyze RNA/RNA and RNA/protein interactions involved in small noncoding RNA-mediated gene regulation networks. In this chapter, we describe and discuss protocols that can be used to measure the dynamics of these interactions, with a particular emphasis on the advantages-and experimental pitfalls-of using the smFRET methodology to study sRNA-based biological systems.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
Host Factor 1 Protein
/
Fluorescence Resonance Energy Transfer
/
RNA, Small Untranslated
/
Fluorescent Dyes
/
Single Molecule Imaging
Language:
En
Year:
2018
Type:
Article