A5082368830- Neurobiology and Insect Physiology Research
- Cellular transport and secretion
- CRISPR and Genetic Engineering
- Retinal Development and Disorders
- Neuroscience and Neuropharmacology Research
- Genetics, Aging, and Longevity in Model Organisms
- Insect symbiosis and bacterial influences
- Ion channel regulation and function
- RNA modifications and cancer
- Chromosomal and Genetic Variations
- Epigenetics and DNA Methylation
- Diet and metabolism studies
- Neuroscience and Neural Engineering
- Lipid Membrane Structure and Behavior
- Mitochondrial Function and Pathology
- Developmental Biology and Gene Regulation
- Metal-Organic Frameworks: Synthesis and Applications
- Cellular Mechanics and Interactions
- Single-cell and spatial transcriptomics
- Cancer-related gene regulation
- Photoreceptor and optogenetics research
- Neural dynamics and brain function
- Genomics and Chromatin Dynamics
- Metalloenzymes and iron-sulfur proteins
- Hippo pathway signaling and YAP/TAZ
Brown University
2019-2025
Allen Institute for Brain Science
2019-2025
Providence College
2022-2025
John Brown University
2019-2024
University of Wisconsin–Madison
2008-2018
Laboratory of Molecular Genetics
2013
Washington University in St. Louis
2003
Abstract We have adapted a bacterial CRISPR RNA/Cas9 system to precisely engineer the Drosophila genome and report that Cas9-mediated genomic modifications are efficiently transmitted through germline. This RNA-guided Cas9 can be rapidly programmed generate targeted alleles for probing gene function in Drosophila.
We and others recently demonstrated that the readily programmable CRISPR/Cas9 system can be used to edit Drosophila genome. However, most applications date have relied on aberrant DNA repair stochastically generate frameshifting indels adoption has been limited by a lack of tools for efficient identification targeted events. Here we report optimized techniques expanded application in through homology-directed (HDR) with double-stranded (dsDNA) donor templates facilitate complex genome...
Multiple stringent confinement strategies should be used whenever possible
Abstract The CRISPR‐Cas9 system has transformed genome engineering of model organisms from possible to practical. can be readily programmed generate sequence‐specific double‐strand breaks that disrupt targeted loci when repaired by error‐prone non‐homologous end joining (NHEJ) or catalyze precise modification through homology‐directed repair (HDR). Here we describe a streamlined approach for rapid and highly efficient the Drosophila via CRISPR‐Cas9‐mediated HDR. In this approach, transgenic...
Drosophila melanogaster is a powerful, long-standing model for metazoan development and gene regulation. We profiled chromatin accessibility in almost 1 million expression half nuclei from overlapping windows spanning the entirety of embryogenesis. Leveraging developmental asynchronicity within embryo collections, we applied deep neural networks to infer age each nucleus, resulting continuous, multimodal views molecular cellular transitions absolute time. identify cell lineages; their...
The CRISPR/Cas9 system has attracted significant attention for its potential to transform genome engineering. We and others have recently shown that the RNA-guided Cas9 nuclease can be employed engineer Drosophila genome, these modifications are efficiently transmitted through germline. A single targeting RNA guide a specific genomic sequence where it induces double-strand breaks that, when imperfectly repaired, yield mutations. also demonstrated 2 RNAs used generate large defined deletions...
Neurons communicate through Ca<sup>2+</sup>-dependent neurotransmitter release at presynaptic active zones (AZs). Neurotransmitter properties play a key role in defining information flow circuits and are tuned during multiple forms of plasticity. Despite their central determining properties, little is known about how Ca<sup>2+</sup> channel levels modulated to calibrate synaptic function. We used CRISPR tag the <i>Drosophila</i> Ca<sub>V</sub>2 Cacophony (Cac) and, males which all Cac...
At presynaptic active zones (AZs), conserved scaffold protein architectures control synaptic vesicle (SV) release by defining the nanoscale distribution and density of voltage-gated Ca 2+ channels (VGCCs). While AZs can potentiate SV in minutes range, we lack an understanding how AZ components VGCCs engage into potentiation. We here establish dynamic, intravital single-molecule imaging endogenously tagged proteins at Drosophila undergoing homeostatic During potentiation, numbers α1 VGCC...
The strength of synaptic connections varies significantly and is a key determinant communication within neural circuits. Mechanistic insight into presynaptic factors that establish modulate neurotransmitter release properties crucial to understanding synapse strength, circuit function, plasticity. We previously identified Drosophila Piccolo-RIM-related Fife, which regulates neurotransmission motor behavior through an unknown mechanism. Here, we demonstrate Fife localizes interacts with RIM...
Post-transcriptional modification of RNA regulates gene expression at multiple levels. ALKBH8 is a tRNA-modifying enzyme that methylates wobble uridines in subset tRNAs to modulate translation. Through methylation tRNA-selenocysteine, promotes selenoprotein synthesis and redox homeostasis. Pathogenic variants have been linked intellectual disability disorders the human population, but role nervous system unknown. vivo studies Drosophila , we show controls oxidative stress brain restrain...
Synaptic heterogeneity is a hallmark of nervous systems that enables complex and adaptable communication in neural circuits. To understand circuit function, it thus critical to determine the factors contribute functional diversity synapses. We investigated contributions voltage-gated calcium channel (VGCC) abundance, spatial organization, subunit composition synapse among between synapses formed by two closely related Drosophila glutamatergic motor neurons with distinct neurotransmitter...
Abstract Background Bridge‐like lipid transfer proteins (BLTPs) mediate bulk transport at membrane contact sites. Mutations in BLTPs are linked to both early‐onset neurodevelopmental and later‐onset neurodegenerative diseases, including movement disorders. The tissue specificity temporal requirements of disease pathogenesis remain poorly understood. Objective objective this study was determine tissue‐specific aging‐dependent roles for VPS13A BLTP2 using Drosophila models. Methods We...
Highlights•In neurons, PDZD8 primarily localizes to ER-late endosome/lysosome membrane contact sites•PDZD8 is required for activity-dependent synaptic growth•PDZD8 promotes bouton formation through an autophagy-dependent mechanism•PDZD8 lysosome maturation and turnover increase autophagic fluxSummaryBuilding connections requires coordinating a host of cellular activities from cell signaling protein turnover, placing high demand on intracellular communication. Membrane sites (MCSs) formed...
Neuronal communication depends on the precisely orchestrated release of neurotransmitter at specialized sites called active zones (AZs). A small number scaffolding and cytoskeletal proteins comprising cytomatrix zone (CAZ) are thought to organize architecture functional properties AZs. The majority CAZ evolutionarily conserved, underscoring fundamental similarities in neurotransmission all synapses. However, core Piccolo Bassoon have long been believed exclusive vertebrates, raising...
A cross-campus, cross-career stage and cross-disciplinary series of discussions at a large public university has produced recommendations for addressing the problems confronting biomedical research community in US.
Asymmetric cell divisions generate sibling cells of distinct fates (‘A’, ‘B’) and constitute a fundamental mechanism that creates cell-type diversity in multicellular organisms. Antagonistic interactions between the Notch pathway intrinsic cell-fate determinant Numb appear to regulate asymmetric flies vertebrates. During these divisions, productive signaling requires sanpodo, which encodes novel transmembrane protein. Here, we demonstrate Drosophila sanpodo plays dual role during —...
SUMMARY Building synaptic connections, which are often far from the soma, requires coordinating a host of cellular activities transcription to protein turnover, placing high demand on intracellular communication. Membrane contact sites (MCSs) formed between organelles have emerged as key signaling hubs for an array activities. We found that endoplasmic reticulum (ER) MCS tethering PDZD8 is required activity-dependent synaptogenesis. sufficient drive ectopic bouton formation through...
Nervous wreck (Nwk) is a conserved F-BAR protein that attenuates synaptic growth and promotes function in Drosophila. In an effort to understand how Nwk carries out its dual roles, we isolated interacting proteins using mass spectrometry. We report interaction between BAR-SH3 sorting nexins, family of membrane-binding implicated diverse intracellular trafficking processes. mammalian cells, nexins induce plasma membrane tubules localize NWK2, consistent with possible functional during the...
We combined cryo-preservation of intact Drosophila larvae and electron tomography with comprehensive segmentation key features to reconstruct the complete ultrastructure a model glutamatergic synapse in near-to-native state. Presynaptically, we detail complex network filaments that connects organizes synaptic vesicles. link complexity this vesicle proximity active zone cytomatrix, consistent these protein structures function together regulate pools. identify net-shaped electron-dense...