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Designed sensors comprising split-firefly luciferase conjugated to tandem poly(ADP-ribose) binding domains allow for the direct solution phase detection of picogram quantities of PAR and for monitoring temporal changes in poly(ADP-ribosyl)ation events in mammalian cells.
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http://dx.doi.org/10.1039/c0cc02229b | DOI Listing |
Bioorg Chem
January 2025
Department of Chemistry, Binghamton University, The State University of New York, 4400 Vestal Parkway East Binghamton, New York 13902, USA. Electronic address:
A modified enzyme fragment complementation assay has been designed and validated as a turn-on biosensor for nucleic acid detection in dilute aqueous solution. The assay is target sequence-agonistic and uses fragments of NanoBiT, the split luciferase reporter enzyme, that are esterified enzymatically at their C-termini to steramers, sterol-linked oligonucleotides. The Drosophila hedgehog autoprocessing domain, DHhC, serves as the self-cleaving enzyme for the NanoBiT-steramer bioconjugations.
View Article and Find Full Text PDFRSC Chem Biol
February 2024
Laboratory of Protein Engineering, Department of Biomedical Engineering and Institute for Complex Molecular Systems, Eindhoven University of Technology Eindhoven The Netherlands
Fast and reliable virus diagnostics is key to prevent the spread of viruses in populations. A hallmark of viruses is the presence of multivalent surface proteins, a property that can be harnessed to control conformational switching in sensor proteins. Here, we introduce a new sensor platform (dark-LUX) for the detection of viral surface proteins consisting of a general bioluminescent framework that can be post-translationally functionalized with separately expressed binding domains.
View Article and Find Full Text PDFFront Genome Ed
March 2022
Genome Center and Department of Biochemistry and Molecular Medicine, University of California, Davis, Davis, CA, United States.
An extensive arsenal of biosensing tools has been developed based on the clustered regularly interspaced short palindromic repeat (CRISPR) platform, including those that detect specific DNA sequences both and in live cells. To date, DNA imaging approaches have traditionally used full fluorescent reporter-based fusion probes. Such "always-on" probes differentiate poorly between bound and unbound probe and are unable to sensitively detect unique copies of a target sequence in individual cells.
View Article and Find Full Text PDFBiosens Bioelectron
February 2021
College of Life Sciences, South-Central University for Nationalities, Wuhan, 430074, PR China. Electronic address:
Detection of dysregulated circulating microRNAs (miRNAs) in human biofluids is a fundamental ability to determine tumor occurrence and metastasis in a minimally invasive fashion. However, the requirements for sophisticated instruments and professional personnel impede the translation of miRNA tests into routine clinical diagnostics, especially for resource-limited regions. Herein, we developed a DNA-guided bioluminescence strategy for the detection of circulating miRNAs.
View Article and Find Full Text PDFACS Sens
July 2020
Department of Biomedical Engineering, Vanderbilt University, Nashville, Tennessee 37235, United States.
Endosomal escape is a critical step in the intracellular delivery of biomacromolecular drugs, but a quantitative, high-throughput study of endosomal-vesicle disruption remains elusive. We designed two genetically encoded split-luciferase turn-on reporter assays that can be measured rapidly in well plates on live cells using a luminometer. Both systems use nonluminescent N-terminal and C-terminal luciferase fragments that can reconstitute a functional luminescent enzyme when they are colocalized by their fusion partners.
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