Probing Nucleic Acid Interactions and Pre-mRNA Splicing by Förster Resonance Energy Transfer (FRET) Microscopy
نویسندگان
چکیده
Förster resonance energy transfer (FRET) microscopy is a powerful technique routinely used to monitor interactions between biomolecules. Here, we focus on the techniques that are used for investigating the structure and interactions of nucleic acids (NAs). We present a brief overview of the most commonly used FRET microscopy techniques, their advantages and drawbacks. We list experimental approaches recently used for either in vitro or in vivo studies. Next, we summarize how FRET contributed to the understanding of pre-mRNA splicing and spliceosome assembly.
منابع مشابه
A FRET-based assay for characterization of alternative splicing events using peptide nucleic acid fluorescence in situ hybridization
We describe a quantitative method for detecting RNA alternative splicing variants that combines in situ hybridization of fluorescently labeled peptide nucleic acid (PNA) probes with confocal microscopy Förster resonance energy transfer (FRET). The use of PNA probes complementary to sequences flanking a given splice junction allows to specifically quantify, within the cell, the RNA isoform gener...
متن کاملDeep-tissue photoacoustic tomography of Förster resonance energy transfer.
Förster resonance energy transfer (FRET) is a distance-dependent process that transfers excited state energy from a donor molecule to an acceptor molecule without the emission of a photon. The FRET rate is determined by the proximity between the donor and the acceptor molecules; it becomes significant only when the proximity is within several nanometers. Therefore, FRET has been applied to visu...
متن کاملSingle-Molecule Pull-Down FRET to Dissect the Mechanisms of Biomolecular Machines.
Spliceosomes are multimegadalton RNA-protein complexes responsible for the faithful removal of noncoding segments (introns) from pre-messenger RNAs (pre-mRNAs), a process critical for the maturation of eukaryotic mRNAs for subsequent translation by the ribosome. Both the spliceosome and ribosome, as well as many other RNA and DNA processing machineries, contain central RNA components that endow...
متن کاملMapping Interactions between mRNA Export Factors in Living Cells
The TREX complex couples nuclear mRNA processing events with subsequent export to the cytoplasm. TREX also acts as a binding platform for the mRNA export receptor Nxf1. The sites of mRNA transcription and processing within the nucleus have been studied extensively. However, little is known about where TREX assembly takes place and where Nxf1 is recruited to TREX to form the export competent mRN...
متن کاملFörster resonance energy transfer photoacoustic microscopy.
Förster, or fluorescence, resonance energy transfer (FRET) provides fluorescence signals sensitive to intra- and inter-molecular distances in the 1 to 10 nm range. Widely applied in the fluorescence imaging environment, FRET enables visualization of physicochemical processes in molecular interactions and conformations. In this paper, we report photoacoustic imaging of FRET, based on nonradiativ...
متن کامل