Thomas A. Kirkland
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View article: In Vitro Psilocybin Synthesis by Co‐Immobilized Enzymes
In Vitro Psilocybin Synthesis by Co‐Immobilized Enzymes Open
Advanced clinical trials investigate the Psilocybe magic mushroom natural product psilocybin as a treatment against major depressive disorder. Currently, synthetic material is used to meet the demand for legitimate pharmaceutical purposes.…
View article: Rapid optimization of protein function in mammalian cells via microbe-independent deep assembly and screening
Rapid optimization of protein function in mammalian cells via microbe-independent deep assembly and screening Open
Random mutagenesis and deep mutational scanning (DMS) are widely used to optimize proteins by oversampling large libraries in microbial cells, selecting cells expressing favorable variants, and sequencing to identify enriched variants. How…
View article: Optimized in vivo multispectral bioluminescent imaging of tumor biology using engineered BRET reporters
Optimized in vivo multispectral bioluminescent imaging of tumor biology using engineered BRET reporters Open
The ability to visualize and track multiple biological processes in vivo in real time is highly desirable. Bioluminescence imaging (BLI) has emerged as an attractive modality for non-invasive cell tracking, with various luciferase reporter…
View article: A Novel Luciferase-Based Reporter Gene Technology for Simultaneous Optical and Radionuclide Imaging of Cells
A Novel Luciferase-Based Reporter Gene Technology for Simultaneous Optical and Radionuclide Imaging of Cells Open
Multimodality reporter gene imaging combines the sensitivity, resolution and translational potential of two or more signals. The approach has not been widely adopted by the animal imaging community, mainly because its utility in this area …
View article: Kinase-Modulated Bioluminescent Indicators Enable Noninvasive Imaging of Drug Activity in the Brain
Kinase-Modulated Bioluminescent Indicators Enable Noninvasive Imaging of Drug Activity in the Brain Open
Aberrant kinase activity contributes to the pathogenesis of brain cancers, neurodegeneration, and neuropsychiatric diseases, but identifying kinase inhibitors that function in the brain is challenging. Drug levels in blood do not predict e…
View article: Kinase-modulated bioluminescent indicators enable noninvasive imaging of drug activity in the brain
Kinase-modulated bioluminescent indicators enable noninvasive imaging of drug activity in the brain Open
Aberrant kinase activity contributes to the pathogenesis of brain cancers, neurodegeneration, and neuropsychiatric diseases, but identifying kinase inhibitors that function in the brain is challenging. Drug levels in blood do not predict e…
View article: Simple, Rapid Chemical Labeling and Screening of Antibodies with Luminescent Peptides
Simple, Rapid Chemical Labeling and Screening of Antibodies with Luminescent Peptides Open
[Image: see text] Sensitive and selective detection assays are essential for the accurate measurement of analytes in both clinical and research laboratories. Immunoassays that rely on nonoverlapping antibodies directed against the same tar…
View article: Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression
Fatal Neurodissemination and SARS-CoV-2 Tropism in K18-hACE2 Mice Is Only Partially Dependent on hACE2 Expression Open
Animal models recapitulating COVID-19 are critical to enhance our understanding of SARS-CoV-2 pathogenesis. Intranasally inoculated transgenic mice expressing human angiotensin-converting enzyme 2 under the cytokeratin 18 promoter (K18-hAC…
View article: In Vitro and In Vivo Analysis of Extracellular Vesicle‐Mediated Metastasis Using a Bright, Red‐Shifted Bioluminescent Reporter Protein (Advanced Genetics 1/03)
In Vitro and In Vivo Analysis of Extracellular Vesicle‐Mediated Metastasis Using a Bright, Red‐Shifted Bioluminescent Reporter Protein (Advanced Genetics 1/03) Open
Extracellular Vesicle Imaging Cancer cell‐derived extracellular vesicles (EVs) promote tumor growth and spread. Studying the distribution of EVs in the body is key to understanding cancer progression and developing therapies. In article nu…
View article: In Vitro and In Vivo Analysis of Extracellular Vesicle‐Mediated Metastasis Using a Bright, Red‐Shifted Bioluminescent Reporter Protein
In Vitro and In Vivo Analysis of Extracellular Vesicle‐Mediated Metastasis Using a Bright, Red‐Shifted Bioluminescent Reporter Protein Open
Cancer cells produce heterogeneous extracellular vesicles (EVs) as mediators of intercellular communication. This study focuses on a novel method to image EV subtypes and their biodistribution in vivo. A red‐shifted bioluminescence resonan…
View article: An Integrated Approach toward NanoBRET Tracers for Analysis of GPCR Ligand Engagement
An Integrated Approach toward NanoBRET Tracers for Analysis of GPCR Ligand Engagement Open
Gaining insight into the pharmacology of ligand engagement with G-protein coupled receptors (GPCRs) under biologically relevant conditions is vital to both drug discovery and basic research. NanoLuc-based bioluminescence resonance energy t…
View article: Small Molecule–Protein Hybrid for Voltage Imaging via Quenching of Bioluminescence
Small Molecule–Protein Hybrid for Voltage Imaging via Quenching of Bioluminescence Open
We report a small-molecule enzyme pair for optical voltage sensing via quenching of bioluminescence. This quenching bioluminescent voltage indicator, or Q-BOLT, pairs the dark absorbing, voltage-sensitive dipicrylamine with membrane-locali…
View article: Streamlined Target Deconvolution Approach Utilizing a Single Photoreactive Chloroalkane Capture Tag
Streamlined Target Deconvolution Approach Utilizing a Single Photoreactive Chloroalkane Capture Tag Open
Identification of physiologically relevant targets for lead compounds emerging from drug discovery screens is often the rate-limiting step toward understanding their mechanism of action and potential for undesired off-target effects. To th…
View article: A Small Molecule – Protein Hybrid for Voltage Imaging via Quenching of Bioluminescence
A Small Molecule – Protein Hybrid for Voltage Imaging via Quenching of Bioluminescence Open
We report a small molecule enzyme pair for optical voltage sensing via quenching of bioluminescence. This Q uenching B ioluminescent V olt age Indicator, or Q-BOLT, pairs the dark absorbing, voltage-sensitive dipicrylamine with membrane-lo…
View article: A Small Molecule – Protein Hybrid for Voltage Imaging via Quenching of Bioluminescence
A Small Molecule – Protein Hybrid for Voltage Imaging via Quenching of Bioluminescence Open
We report a small molecule enzyme pair for optical voltage sensing via quenching of bioluminescence. This Quenching Bioluminescent Voltage Indicator, or Q-BOLT, pairs the dark absorbing, voltage-sensitive dipicrylamine with membrane-locali…
View article: 5,5-Dialkylluciferins are thermal stable substrates for bioluminescence-based detection systems
5,5-Dialkylluciferins are thermal stable substrates for bioluminescence-based detection systems Open
Firefly luciferase-based ATP detection assays are frequently used as a sensitive, cost-efficient method for monitoring hygiene in many industrial settings. Solutions of detection reagent, containing a mixture of a substrate and luciferase …
View article: NanoBiT System and Hydrofurimazine for Optimized Detection of Viral Infection in Mice—A Novel in Vivo Imaging Platform
NanoBiT System and Hydrofurimazine for Optimized Detection of Viral Infection in Mice—A Novel in Vivo Imaging Platform Open
Reporter genes are used to visualize intracellular biological phenomena, including viral infection. Here we demonstrate bioluminescent imaging of viral infection using the NanoBiT system in combination with intraperitoneal injection of a f…
View article: Abstract 1957: Novel substrates for NanoLuc luciferase with improved brightness and signal duration for bioluminescence imaging<i>in vivo</i>
Abstract 1957: Novel substrates for NanoLuc luciferase with improved brightness and signal duration for bioluminescence imaging<i>in vivo</i> Open
NanoLuc® luciferase, and its substrate furimazine, are becoming a preferred bioluminescent system in cellular and biochemical assays due to its brightness and glow-type kinetics. The construct Antares, made by fusing NanoLuc to the cyan-ex…
View article: Homogeneous Assay for Target Engagement Utilizing Bioluminescent Thermal Shift
Homogeneous Assay for Target Engagement Utilizing Bioluminescent Thermal Shift Open
Protein thermal shift assays (TSAs) provide a means for characterizing target engagement through ligand-induced thermal stabilization. Although these assays are widely utilized for screening libraries and validating hits in drug discovery …
View article: Quantitative, Wide-Spectrum Kinase Profiling in Live Cells for Assessing the Effect of Cellular ATP on Target Engagement
Quantitative, Wide-Spectrum Kinase Profiling in Live Cells for Assessing the Effect of Cellular ATP on Target Engagement Open
For kinase inhibitors, intracellular target selectivity is fundamental to pharmacological mechanism. Although a number of acellular techniques have been developed to measure kinase binding or enzymatic inhibition, such approaches can fail …