Adeno-associated viral vectors for functional intravenous gene transfer throughout the non-human primate brain
By onCrossing the blood–brain barrier in primates is a major obstacle for gene delivery to the brain. Adeno-associated viruses (AAVs) promise robust, non-invasive gene delivery from the bloodstream to the brain. However, unlike in rodents, few neurotropic AAVs efficiently cross the blood–brain barrier in non-human primates. Here we report on AAV.CAP-Mac, an engineered variant identified by screening in adult marmosets and newborn macaques, which has improved delivery efficiency in the brains of multiple non-human primate species: marmoset, rhesus macaque and green monkey. CAP-Mac is neuron biased in infant Old World primates, exhibits broad tropism in adult rhesus macaques and is vasculature biased in adult marmosets. We demonstrate applications of a single, intravenous dose of CAP-Mac to deliver functional GCaMP for ex vivo calcium imaging across multiple brain areas, or a cocktail of fluorescent reporters for Brainbow-like labelling throughout the macaque brain, circumventing the need for germline manipulations in Old World primates. As such, CAP-Mac is shown to have potential for non-invasive systemic gene transfer in the brains of non-human primates.
pCAG-MBP-Foldon-ATG9 (801-839)
By onPlasmid: To make ATG9 C-terminal tail trimer for expression in mammalian cells.
3D FIBSEM CLEM
By onThis protocol details the procedure of Correlative Light Microscopy and Electron Microscopy (CLEM) with 3D Focus Ion Beam Scanning Electron Microscopy (FIBSEM) technique in Zeiss Crossbeam 550 FIBSEM system.
Structure of human ULK1C:PI3KC3-C1 supercomplex
By onStructure of human ULK1C:PI3KC3-C1 supercomplex
Th-cre;Ercc1-/fl mice
By onDopaminergic neurons may experience elevated DNA damage and potential senescence, suggesting implications for neurodegenerative disorders.
Differential response of α-synuclein expression to bacterial ligands and metabolites in mouse enteroendocrine cells
By onDataset for manuscript " α-synuclein expression in response to bacterial ligands and metabolites in gut enteroendocrine cells". Tabs in excel file are title with the figure number.
Structural and biochemical insights into lipid transport by VPS13 proteins
By onVPS13 proteins are proposed to function at contact sites between organelles as bridges for lipids to move directionally and in bulk between organellar membranes. VPS13s are anchored between membranes via interactions with receptors, including both peripheral and integral membrane proteins. Here we present the crystal structure of VPS13s adaptor binding domain (VAB) complexed with a Pro-X-Pro peptide recognition motif present in one such receptor, the integral membrane protein Mcp1p, and show biochemically that other Pro-X-Pro motifs bind the VAB in the same site. We further demonstrate that Mcp1p and another integral membrane protein that interacts directly with human VPS13A, XK, are scramblases. This finding supports an emerging paradigm of a partnership between bulk lipid transport proteins and scramblases. Scramblases can re-equilibrate lipids between membrane leaflets as lipids are removed from or inserted into the cytosolic leaflet of donor and acceptor organelles, respectively, in the course of protein-mediated transport.
Transfection and protein over-expression
By onThis protocol provides the details for how to transfect and harvest 3X-FLAG-SHIP164Δ901-1099
Fast Scan Cyclic Voltammetry (FSCV) in Mouse Brain Slices
By onThis protocol describes the method of fast scan cyclic voltammetry (FSCV) for the application of measuring dopamine transients in mouse brain slices. Within the protocol are sections describing the manufacture of microelectrodes, preparation of acute brain slices, fiber calibration, and measuring stimulated dopamine release.
Live-cell imaging; mitochondrial reactive oxygen species
By onThis protocol presents instructions for assessing mitochondrial souced reactive oxygen species using MitoTracker® Red CM-H2XRos dye.
Simulations predict differing phase responses to excitation vs. inhibition intheta-resonant pyramidal neurons
By onThis research highlights how cortical neurons respond differently to oscillatory inputs, impacting neuronal responses to network state shifts.
Expression and purification of PI3KC3-C1 complex
By onProtocol describing expression and purification of PI3KC3-C1 complex.
TBK1 knockdown and rescue in Hela-M cells
By onThis protocol transiently depletes TBK1 from HeLa cells and re-introduces a tagged TBK1 along with other relevant components of mitophagy.
Synapsin E-domain is essential for α-synuclein function
By onSynucleins and synapsins are thought to cooperate to regulate synaptic vesicle recycling, but the mechanism is not known. Here we identify the synapsin E-domain as an essential binding partner of α-syn, enabling the function of α-syn at the synapse.
Genome-wide determinants of mortality and clinical progression in Parkinson’s disease – Summary statistics
By onSummary statistics from "Genome-wide determinants of mortality and clinical progression in Parkinson’s disease."
Primary data associated with the manuscript “Genome-wide screen reveals Rab12 GTPase as 2 a critical activator of Parkinson’s disease-linked LRRK2 kinase” (3/3)
By onPrimary data associated with the manuscript, "Genome-wide screen reveals Rab12 GTPase as 2 a critical activator of Parkinson’s disease-linked LRRK2 kinase."
AAVS1 Knock-in
By onThis protocol describes the standard procedure to knock-in constructs to the AAVS1 safe harbor locus in hPSCs.
