ArticleBiotechnology progress
Rational design of novel red-shifted BRET pairs: Platforms for real-time single-chain protease biosensors.
Article in Biotechnology progress. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 44 citations in OpenAlex.
- Using a bioluminescence resonance energy transfer caspase biosensor to study caspase-3 cleavage site specificity.Bioscience reports · 2026Article
- Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer.Scientific reports · 2021Article
- Development of a Bio-Layer Interferometry-Based Protease Assay Using HIV-1 Protease as a Model.Viruses · 2021Article
- Interrogating Cellular Communication in Cancer with Genetically Encoded Imaging Reporters.Radiology. Imaging cancer · 2020Review
- Bright Bioluminescent BRET Sensor Proteins for Measuring Intracellular Caspase Activity.ACS sensors · 2017Article
- Methods used to study the oligomeric structure of G-protein-coupled receptors.Bioscience reports · 2017Review
- Redshifted Cherenkov Radiation for in vivo Imaging: Coupling Cherenkov Radiation Energy Transfer to multiple Förster Resonance Energy Transfers.Scientific reports · 2017Article
- In Vivo Analysis of Protein-Protein Interactions with Bioluminescence Resonance Energy Transfer (BRET): Progress and Prospects.International journal of molecular sciences · 2016Review
- Fabrication of molecular tension probes.MethodsX · 2016Article
- Fluorophore-NanoLuc BRET Reporters Enable Sensitive In Vivo Optical Imaging and Flow Cytometry for Monitoring Tumorigenesis.Cancer research · 2015Article
- Small Molecule Active Site Directed Tools for Studying Human Caspases.Chemical reviews · 2015Review
- High Throughput Screening and Selection Methods for Directed Enzyme Evolution.Industrial & engineering chemistry research · 2015Review
- Bioluminescence: a versatile technique for imaging cellular and molecular features.MedChemComm · 2014Article
- Making the brain glow: in vivo bioluminescence imaging to study neurodegeneration.Molecular neurobiology · 2013Review
- Bioluminescence resonance energy transfer methods to study G protein-coupled receptor-receptor tyrosine kinase heteroreceptor complexes.Methods in cell biology · 2013Article
- Biomarkers and molecular probes for cell death imaging and targeted therapeutics.Bioconjugate chemistry · 2012Review
- Design of "smart" probes for optical imaging of apoptosis.American journal of nuclear medicine and molecular imaging · 2011Article
- Genetically encodable fluorescent biosensors for tracking signaling dynamics in living cells.Chemical reviews · 2011Review
- Cerenkov radiation energy transfer (CRET) imaging: a novel method for optical imaging of PET isotopes in biological systems.PloS one · 2010Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Bioluminescence resonance energy transfer (BRET) systems to date have been dominated by use of blue-green Renilla luciferase (Rluc) as the energy donor. Although effective in many cases, the expense and unfavorable biochemical attributes of the substrate (phenylcoelenterazine) limit utility of Rluc-based BRET systems. Herein we report a series of novel BRET pairs based on luciferases that utilize D-luciferin, resulting in red-shifted photonic outputs, favorable biochemical attributes, and increased efficacy. We developed a modified Förster equation to predict optimal BRET luciferase donor-fluorophore pairs and identified tdTomato as the optimal red fluorophore acceptor for click beetle green luciferase (CBG). A prototypical single-chain protease biosensor, capable of reporting on executioner caspase activity in live cells and in real-time, was generated by inserting a DEVD linker between CBG and tdTomato and validated in vitro with recombinant caspases and in cellulo with apoptosis-sensitive and -resistant cell lines. High signal-to-noise ratios ( approximately 33) and Z' factors (0.85) were observed in live cell longitudinal studies, sufficient for high-throughput screening. Thus, we illustrate a general methodology for the rational design of new BRET systems and provide a novel single-chain BRET protease biosensor that is long lived, red-shifted, and utilizes D-luciferin.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.