A5008140261- Cannabis and Cannabinoid Research
- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- Receptor Mechanisms and Signaling
- Neuroscience and Neuropharmacology Research
- Cellular transport and secretion
- Pancreatic function and diabetes
- Virus-based gene therapy research
- CRISPR and Genetic Engineering
- Pharmacological Receptor Mechanisms and Effects
- Endoplasmic Reticulum Stress and Disease
- Lipid Membrane Structure and Behavior
- Calcium signaling and nucleotide metabolism
- Biotin and Related Studies
- Protein Structure and Dynamics
- Monoclonal and Polyclonal Antibodies Research
- Advanced Electron Microscopy Techniques and Applications
- Neurotransmitter Receptor Influence on Behavior
- Retinoids in leukemia and cellular processes
- Microtubule and mitosis dynamics
- Neurological disorders and treatments
- Ion channel regulation and function
- Parkinson's Disease Mechanisms and Treatments
- Phagocytosis and Immune Regulation
- RNA Research and Splicing
AstraZeneca (United Kingdom)
2018-2024
University of Auckland
2014-2019
University of Cambridge
2016-2018
Background and Purpose CB 1 receptor signalling is canonically mediated through inhibitory Gα i proteins, but occurs other G proteins under some circumstances, s being the most characterized secondary pathway. Determinants of this switch identified to date include blockade, /D 2 co‐stimulation, agonist class cell background. Hence, we examined effects number different ligands on signalling. Experimental Approach receptors were expressed in HEK cells at levels, for cAMP by real‐time BRET...
Abstract Recently, lipid nanoparticles (LNPs) have attracted attention due to their emergent use for COVID‐19 mRNA vaccines. The success of LNPs can be attributed ionizable lipids, which enable functional intracellular delivery. Previously, the authors established an automated high‐throughput platform screen lipids and identified that generated using this technique show comparable or increased delivery in vitro as compared prepared traditional microfluidics techniques. In study, choose one...
Despite recent advances in the field of mRNA therapy, lack safe and efficacious delivery vehicles with pharmaceutically developable properties remains a major limitation. Here, we describe systematic optimisation lipid-peptide nanocomplexes for two murine cancer cell types, B16-F10 melanoma CT26 colon carcinoma as well NCI-H358 human lung bronchoalveolar cells. Different combinations lipids peptides were screened from an original nanocomplex formulation improved luciferase transfection vitro...
Abstract Poor understanding of intracellular delivery and targeting hinders development nucleic acid‐based therapeutics transported by nanoparticles. Utilizing a siRNA‐targeting small molecule profiling approach with advanced imaging machine learning biological insights is generated into the mechanism lipid nanoparticle (MC3‐LNP) mRNA. This workflow termed Advanced Cellular Endocytic for Intracellular Delivery (ACE‐ID). A cell‐based assay perturbation 178 targets relevant to trafficking used...
Consensus protein design is a rapid and reliable technique for the improvement of stability, which relies on use homologous sequences. To enhance stability fibronectin type III (FN3) domain, consensus was employed using an alignment 2123 The resulting FN3 FN3con, has unprecedented with melting temperature >100°C, ΔG(D-N) 15.5 kcal mol(-1) greatly reduced unfolding rate compared wild-type. determine underlying molecular basis X-ray crystal structure FN3con determined to 2.0 Å other domains...
The microtubule motor dynein plays a key role in cellular organization. However, little is known about how dynein’s biosynthesis, assembly, and functional diversity are orchestrated. To address this issue, we have conducted an arrayed CRISPR loss-of-function screen human cells using the distribution of dynein-tethered peroxisomes early endosomes as readouts. From genome-wide gRNA library, 195 validated hits were recovered parsed into those impacting multiple cargoes whose effects restricted...
Eukaryotic cells use conserved multisubunit membrane tethering complexes, including CORVET (class C core vacuole/endosome tethering) and HOPS (homotypic fusion vacuole protein sorting), to control the of endomembranes. These complexes have been extensively studied in yeast, but date there far fewer studies metazoan HOPS. Both these comprise six subunits: a common four-subunit two unique subunits. Once assembled, function recognise specific endosomal markers facilitate SNARE-mediated fusion....
Multi-subunit tethering complexes control membrane fusion events in eukaryotic cells. Class C core vacuole/endosome (CORVET) and homotypic vacuole protein sorting (HOPS) are two such complexes, both containing the Sec1/Munc18 subunit VPS33A. Metazoans additionally possess VPS33B, which has considerable sequence similarity to VPS33A but does not integrate into CORVET or HOPS instead stably interacts with VIPAR. It been recently suggested that VPS33B VIPAR comprise subunits of a novel...
The favorable biophysical attributes of non-antibody scaffolds make them attractive alternatives to monoclonal antibodies. However, due the well-known stability-function trade-off, these gains tend be marginal after functional selection. A notable example is fibronectin Type III (FN3) domain, FNfn10, which has been previously evolved bind lysozyme with 1 pM affinity (FNfn10-α-lys), but suffers from poor thermodynamic and kinetic stability. To explore this compromise further, we grafted...
Modified messenger RNAs (mRNAs) hold great potential as therapeutics by using the body's own processes for protein production. However, a key challenge is efficient delivery of therapeutic mRNA to cell cytosol and productive translation. Lipid nanoparticles (LNPs) are most clinically advanced system nucleic acid delivery; however, relatively narrow index makes them unsuitable many applications. A obstacle development more potent LNPs limited mechanistic understanding interaction with cells....
Abstract G protein-coupled receptors (GPCRs) are important therapeutic targets and therefore extensively studied. Like most transmembrane proteins, there has been considerable difficulty in developing reliable specific antibodies for them. To overcome this, epitope tags often used to facilitate antibody recognition studies on fundamental receptor signalling trafficking. In our study of cannabinoid CB 1 /dopamine D 2 interactions we sought generate HEK293 cells expressing FLAG-tagged use...
ABSTRACT Multi-subunit tethering complexes control membrane fusion events in eukaryotic cells. CORVET and HOPS are two such multi-subunit complexes, both containing the Sec1/Munc18 protein subunit VPS33A. Metazoans additionally possess VPS33B, which has considerable sequence similarity to VPS33A but does not integrate into or instead stably interacts with VIPAR. It been recently suggested that VPS33B VIPAR comprise subunits of a novel complex (named ‘CHEVI’), analogous configuration HOPS. We...
ABSTRACT Eukaryotic cells use conserved multisubunit membrane tethering complexes, including CORVET and HOPS, to control the fusion of endomembranes. These complexes have been extensively studied in yeast, but date there far fewer studies metazoan HOPS. Both these comprise six subunits: a common four-subunit core two unique subunits. Once assembled, function recognise specific endosomal markers facilitate SNARE-mediated fusion. promotes homotypic early endosomes, while HOPS lysosomes late...