ArticleMolecular biology of the cell2011
Development of an optimized backbone of FRET biosensors for kinases and GTPases.
Article in Molecular biology of the cell, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 334 papers.
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Who cites it
334 citing papers in PubMed.
- Evaluating and comparing the signalling and functional landscape of hiPSC-CMs derived from patients with dilated cardiomyopathy.npj biomedical innovations · 2026Article
- Noninvasive methods to monitor dynamic single-cell events.Trends in cell biology · 2026Review
- Red-to-Near-Infrared Fluorescence Resonance Energy Transfer Reporters for Dual Imaging Applications.Small methods · 2026Article
- Design and optimization of a kinase-controlled allosteric switch.Nature methods · 2026Article
- The working lives of neuronal 5-HTRSC chemical biology · 2026Review
- Combining Fret and Super-Resolution Microscopy Reveals Kinase Activation and Mitochondrial Activity at the Nanoscale.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- ERK builds a population of short-lived nascent adhesions that produce persistent edge protrusion and cell migration.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- A Constellation of Fluorescent Biosensors to Illuminate the Galaxy of Protein Kinases.Chembiochem : a European journal of chemical biology · 2026Review
- Illuminating compartmentalized AMPK signaling in single cells.The Biochemical journal · 2026Review
- Directly adopting inverse biosensors to image live cell enzyme activities in nanodomains.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- ZO-1 shuttles between apical junctional complexes and podosomes by riding ERK activation waves.Nature communications · 2026Article
- Localized heat induces ERK activation and signal propagation in solid tumors.Scientific reports · 2026Article
- Signal control during tissue regeneration in adult animals.Nature reviews. Molecular cell biology · 2026Review
- Real-Time Visualization of Isoform-Specific RAF-KRAS Interactions in Living Cells Using FRET-BRET Hybrid Biosensors.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Traction force transmission via bioactive matrix hydrogel promotes epithelial collective migration mediated by integrin.Scientific reports · 2026Article
- Sensitive and Selective Next-Generation FRET-based PKA Biosensors.bioRxiv : the preprint server for biology · 2026Article
- Robust calibration and quantification of FRET signals using multiplexed biosensor barcoding.iScience · 2025Article
- ERK builds a population of short-lived nascent adhesions that produce persistent edge protrusion and cell migration.bioRxiv : the preprint server for biology · 2025Article
- Article
- Two novel red-FRET ERK bosensors in the 670-720 nm range.Journal of biological engineering · 2025Article
274 more citing papers are in PubMed but not listed here.
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Biosensors based on the principle of Förster (or fluorescence) resonance energy transfer (FRET) have shed new light on the spatiotemporal dynamics of signaling molecules. Among them, intramolecular FRET biosensors have been increasingly used due to their high sensitivity and user-friendliness. Time-consuming optimizations by trial and error, however, obstructed the development of intramolecular FRET biosensors. Here we report an optimized backbone for rapid development of highly sensitive intramolecular FRET biosensors. The key concept is to exclude the "orientation-dependent" FRET and to render the biosensors completely "distance-dependent" with a long, flexible linker. We optimized a pair of fluorescent proteins for distance-dependent biosensors, and then developed a long, flexible linker ranging from 116 to 244 amino acids in length, which reduced the basal FRET signal and thereby increased the gain of the FRET biosensors. Computational simulations provided insight into the mechanisms by which this optimized system was the rational strategy for intramolecular FRET biosensors. With this backbone system, we improved previously reported FRET biosensors of PKA, ERK, JNK, EGFR/Abl, Ras, and Rac1. Furthermore, this backbone enabled us to develop novel FRET biosensors for several kinases of RSK, S6K, Akt, and PKC and to perform quantitative evaluation of kinase inhibitors in living cells.
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