Ratiometric fluorescence imaging of dual bio-molecular events in single living cells using a new FRET pair mVenus/mKOκ-based biosensor and a single fluorescent protein biosensor

Su, Ting; Luo, Qingming

doi:10.1016/j.bios.2011.10.034

Cited by 20 publications

(9 citation statements)

References 31 publications

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“…360 These spectrally compatible probes had minimal cross-talk and were shown to exhibit great spatiotemporal precision. As proof of concept, simultaneous imaging of Src/Ca 2+ signaling and glutathione redox potential were measured, revealing that the epidermal growth factor-induced Src signaling is negatively regulated by H 2 O 2 .…”

Section: Fluorescence Imagingmentioning

confidence: 99%

Single Cell Optical Imaging and Spectroscopy

et al. 2013

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Section: Fluorescence Imagingmentioning

confidence: 99%

Single Cell Optical Imaging and Spectroscopy

et al. 2013

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“…A FRET biosensor based on the mKO/mKate FRET pair was already developed by Su et al, who used mKOκ and mLumin (mKate-S158A) as the donor and acceptor, respectively. 11,53 The authors used the classical design; that is, the ligand and sensor domains were sandwiched with mKOκ and mLumin. The response of such an mKOκ/mLumin-based biosensor for Src was only marginal.…”

Section: ■ Discussionmentioning

confidence: 99%

Booster, a Red-Shifted Genetically Encoded Förster Resonance Energy Transfer (FRET) Biosensor Compatible with Cyan Fluorescent Protein/Yellow Fluorescent Protein-Based FRET Biosensors and Blue Light-Responsive Optogenetic Tools

2020

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Genetically encoded Forster resonance energy transfer (FRET)based biosensors have been developed for the visualization of signaling molecule activities. Currently, most of them are comprised of cyan and yellow fluorescent proteins (CFP and YFP), precluding the use of multiple FRET biosensors within a single cell. Moreover, the FRET biosensors based on CFP and YFP are incompatible with the optogenetic tools that operate at blue light. To overcome these problems, here, we have developed FRET biosensors with red-shifted excitation and emission wavelengths. We chose mKOκ and mKate2 as the favorable donor and acceptor pair by calculating the Forster distance. By optimizing the order of fluorescent proteins and modulatory domains of the FRET biosensors, we developed a FRET biosensor backbone named "Booster". The performance of the protein kinase A (PKA) biosensor based on the Booster backbone (Booster-PKA) was comparable to that of AKAR3EV, a previously developed FRET biosensor comprising CFP and YFP. For the proof of concept, we first showed simultaneous monitoring of activities of two protein kinases with Booster-PKA and ERK FRET biosensors based on CFP and YFP. Second, we showed monitoring of PKA activation by Beggiatoa photoactivated adenylyl cyclase, an optogenetic generator of cyclic AMP. Finally, we presented PKA activity in living tissues of transgenic mice expressing Booster-PKA. Collectively, the results demonstrate the effectiveness and versatility of Booster biosensors as an imaging tool in vitro and in vivo.

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“…Optimizing such approaches, small-angle X-ray scattering for developing calcium sensors and fluorescence resonance energy transfer-probes for kinase sensing are being cited as best biosensors methods in modern physiology (Oldach and Zhang, 2014 ). By this way, several microbial and cell organelle-based biosensors were developed with specific targets (Su et al, 2012 ). As explained earlier, electrochemical, electromechanical, and optical biosensors are developed for detecting miRNA much more efficiently than other molecular techniques (Johnson and Mutharasan, 2014 ).…”

Section: Genetically Encoded or Synthetic Fluorescent Biosensorsmentioning

confidence: 99%

Recent Advances in Biosensor Technology for Potential Applications – An Overview

Vigneshvar

Sudhakumari

Senthilkumaran

et al. 2016

Front. Bioeng. Biotechnol.

474

237

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Imperative utilization of biosensors has acquired paramount importance in the field of drug discovery, biomedicine, food safety standards, defense, security, and environmental monitoring. This has led to the invention of precise and powerful analytical tools using biological sensing element as biosensor. Glucometers utilizing the strategy of electrochemical detection of oxygen or hydrogen peroxide using immobilized glucose oxidase electrode seeded the discovery of biosensors. Recent advances in biological techniques and instrumentation involving fluorescence tag to nanomaterials have increased the sensitive limit of biosensors. Use of aptamers or nucleotides, affibodies, peptide arrays, and molecule imprinted polymers provide tools to develop innovative biosensors over classical methods. Integrated approaches provided a better perspective for developing specific and sensitive biosensors with high regenerative potentials. Various biosensors ranging from nanomaterials, polymers to microbes have wider potential applications. It is quite important to integrate multifaceted approaches to design biosensors that have the potential for diverse usage. In light of this, this review provides an overview of different types of biosensors being used ranging from electrochemical, fluorescence tagged, nanomaterials, silica or quartz, and microbes for various biomedical and environmental applications with future outlook of biosensor technology.

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Ratiometric fluorescence imaging of dual bio-molecular events in single living cells using a new FRET pair mVenus/mKOκ-based biosensor and a single fluorescent protein biosensor

Cited by 20 publications

References 31 publications

Single Cell Optical Imaging and Spectroscopy

Single Cell Optical Imaging and Spectroscopy

Booster, a Red-Shifted Genetically Encoded Förster Resonance Energy Transfer (FRET) Biosensor Compatible with Cyan Fluorescent Protein/Yellow Fluorescent Protein-Based FRET Biosensors and Blue Light-Responsive Optogenetic Tools

Recent Advances in Biosensor Technology for Potential Applications – An Overview

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