A5100650736- Advanced Electron Microscopy Techniques and Applications
- Electron and X-Ray Spectroscopy Techniques
- Bacteriophages and microbial interactions
- RNA and protein synthesis mechanisms
- RNA modifications and cancer
- Single-cell and spatial transcriptomics
- Advanced Biosensing Techniques and Applications
- RNA Interference and Gene Delivery
- Viral Infections and Immunology Research
- Advanced biosensing and bioanalysis techniques
- Receptor Mechanisms and Signaling
- Macrophage Migration Inhibitory Factor
- Photosynthetic Processes and Mechanisms
- Protein Tyrosine Phosphatases
- Neuropeptides and Animal Physiology
- Respiratory viral infections research
- Advanced Fluorescence Microscopy Techniques
- Force Microscopy Techniques and Applications
- Digital Holography and Microscopy
- PI3K/AKT/mTOR signaling in cancer
- Congenital heart defects research
- Biomedical Ethics and Regulation
- Pharmacological Receptor Mechanisms and Effects
- Herpesvirus Infections and Treatments
- Microfluidic and Bio-sensing Technologies
University of California, San Francisco
2025
University of Wisconsin–Madison
2021-2024
Blueprint Medicines (United States)
2012-2021
Akoya Biosciences (United States)
2019-2020
Oregon State University
2018
UCLA Health
2015
Ernest Gallo Clinic and Research Center
2012
Institute of Infection and Immunity
2007
Amgen (United States)
2007
University of California, Berkeley
2000-2004
Imaging large fields of view while preserving high-resolution structural information remains a challenge in low-dose cryo-electron tomography. Here we present robust tools for montage parallel array cryo-tomography (MPACT) tailored vitrified specimens. The combination correlative cryo-fluorescence microscopy, focused-ion-beam milling, substrate micropatterning, and MPACT supports studies that contextually define the three-dimensional architecture cells. To further extend flexibility MPACT,...
Abstract Large library docking of tangible molecules has revealed potent ligands across many targets. While make-on-demand libraries now exceed 75 billion enumerated molecules, their synthetic routes are dominated by a few reaction types, reducing diversity and inevitably leaving interesting bioactive-like chemotypes unexplored. Here, we investigate the large-scale enumeration targeted isoquinuclidines. These “natural-product-like” rare in current functionally congested, making them as...
ADVERTISEMENT RETURN TO ISSUEPREVPerspectiveNEXTConformation-Specific Effects of Raf Kinase InhibitorsMiniperspectiveXiaolun Wang*† and Joseph Kim‡View Author Information† Takeda California, 10410 Science Center Drive, San Diego, California 92121, United States‡ Blueprint Medicines, 215 First Street, Cambridge, Massachusetts 02142, States*Phone: (858) 731-8983. E-mail: [email protected]Cite this: J. Med. Chem. 2012, 55, 17, 7332–7341Publication Date (Web):July 18, 2012Publication History...
Peptide nucleic acids have emerged over the past two decades as a promising class of acid mimics because their strong binding affinity and sequence selectivity toward DNA RNA, resistance to enzymatic degradation by proteases nucleases. While they been shown be effective in regulation gene expression vitro, small extent vivo, full potential for molecular therapy has not yet fully realized due poor cellular uptake. Herein, we report development cell-permeable, guanidine-based peptide targeting...
Abstract Imaging large fields of view while preserving high-resolution structural information remains a challenge in low-dose cryo-electron tomography. Here, we present robust tools for montage electron tomography tailored vitrified specimens. The integration correlative cryo-fluorescence microscopy, focused-ion beam milling, and micropatterning produces contextual three-dimensional architecture cells. Montage tilt series may be processed their entirety or as individual tiles suitable...
Whole-cell cryo-electron tomography (cryo-ET) is a powerful technology that used to produce nanometer-level resolution structures of macromolecules present in the cellular context and preserved near-native frozen-hydrated state. However, there are challenges associated with culturing and/or adhering cells onto TEM grids manner suitable for while retaining their physiological Here, detailed step-by-step protocol presented on use micropatterning direct promote eukaryotic cell growth grids....
An in vitro selection procedure was used to select RNase P ribozyme variants that efficiently cleaved the sequence of mRNA encoding thymidine kinase herpes simplex virus 1. Of 45 selected sequenced, 25 ribozymes carried a common mutation at nucleotides 224 and 225 catalytic RNA from Escherichia coli (G(224)G(225) --> AA). These exhibited least 10 times higher cleavage efficiency (k(cat)/K(m)) than derived wild type ribozyme. Our results suggest mutated A(224)A(225) are close proximity...
Abstract Cellular neurobiology has benefited from recent advances in the field of cryo-electron tomography (cryo-ET). Numerous structural and ultrastructural insights have been obtained plunge-frozen primary neurons cultured on electron microscopy grids. With most having derived rodent sources, we sought to expand breadth sample availability by using 3rd instar Drosophila melanogaster larval brains. Ultrastructural abnormalities were encountered while establishing this model system for...
(<i>S,R</i>)-3-Phenyl-4,5-dihydro-5-isoxasole acetic acid (VGX-1027) is an isoxazole compound that exhibits various immunomodulatory properties. The capacity of VGX-1027 to prevent interleukin (IL)-1β plus interferon-γ-induced pancreatic islet death in vitro prompted us evaluate its effects on the development autoimmune diabetes preclinical models human type 1 mellitus (T1D). Administration NOD mice with spontaneous or accelerated forms induced either by injection cyclophosphamide transfer...
External guide sequences (EGSs) are small RNA molecules that bind to a target mRNA, form complex resembling the structure of tRNA, and render mRNA susceptible hydrolysis by RNase P, tRNA processing enzyme. An in vitro selection procedure was used select EGSs direct human P cleave encoding thymidine kinase (TK) herpes simplex virus 1. One selected EGSs, TK17, at least 35 times more active directing cleaving TK mRNAin than EGS derived from natural sequence. when with sequence, resembles...
Abstract Respiratory syncytial virus (RSV) is an enveloped, filamentous, negative-strand RNA that causes significant respiratory illness worldwide. RSV vaccines are available, however there still need for research to support the development of and therapeutics against related Mononegavirales viruses. Individual virions vary in size, with average diameter ~130 nm ranging from ~500 over 10 µm length. Though general arrangement structural proteins known, we use cryo-electron tomography...
Cryo-electron microscopy and single particle averaging (SPA) has been used to determine the structure of numerous purified proteins resolutions less than three Angstroms [1].SPA can also be applied larger biological assemblies including icosahedral helical virions.In fact, large size global symmetry these particles aid in obtaining high-resolution averages [2,3].However, many viruses other are heterogenous lack true making them unsuitable for SPA.Three-dimensional structures targets obtained...
By using an in vitro selection procedure, we have previously isolated RNase P ribozyme variants that efficiently cleave mRNA sequence vitro. In this study, a variant was used to target the overlapping region of mRNAs encoding human cytomegalovirus (HCMV) major transcription regulatory proteins IE1 and IE2. The is about 90 times more efficient cleaving IE1/IE2 than derived from wild type ribozyme. Our results provide first direct evidence point mutation at nucleotide position 80 catalytic RNA...
Abstract Characterizing the complexities of tumor microenvironment is elemental to understanding disease mechanisms. The spatial relationships between infiltrating immune cells and remodeling cellular matrix widely recognized as a key component defining heterogeneity. Current methodologies for analyzing dimension in tissues, like traditional immunofluorescence (IF) immunohistochemistry (IHC), are limited few parameters at time, restricting scope identifiable cells. Conversely, single-cell...
ABSTRACT Whole-cell cryo-electron tomography (cryo-ET) is a powerful technique that can provide nanometer-level resolution of biological structures within the cellular context and in near-native frozen-hydrated state. It remains challenge to culture or adhere cells on TEM grids manner suitable for while preserving physiological state cells. Here, we demonstrate versatility micropatterning direct promote growth both cultured primary eukaryotic grids. We show compatible with be used enhance...
Abstract Respiratory syncytial virus (RSV) is a significant cause of respiratory illness in young children and adults worldwide. There currently no vaccine or targeted antiviral for RSV. RSV an enveloped, filamentous, negative-strand RNA virus. Individual virions vary both diameter length, with average ∼130 nm ranging from ∼500 to over 10 μm length. The matrix (M) protein peripherally associated the interior viral membrane. Though general arrangement structural proteins within virion known,...
Engineered RNase P ribozymes are promising gene-targeting agents that can be used in both basic research and clinical applications. We have previously selected ribozyme variants for their activity cleaving an mRNA substrate from a pool of containing randomized sequences. In this study, one the was to target encoding thymidine kinase (TK) herpes simplex virus 1 (HSV-1). The variant exhibited enhanced cleavage binding at least 30 times more efficient TK vitro than derived wild type sequence....
Abstract The tumor microenvironment (TME) comprises a multitude of cell types that collectively create an immunosuppressive environment, enabling growth. Understanding the spatial organization these tissues requires technology can identify presence multiple markers and correlate it to their specific location within same tissue. Characteristics TME including infiltrating immune cells, angiogenesis other nonmalignant cells influence important phenomena like drug delivery, treatment...
Whole-cell cryo-electron tomography (cryo-ET) is a powerful technology that used to produce nanometer-level resolution structures of macromolecules present in the cellular context and preserved near-native frozen-hydrated state. However, there are challenges associated with culturing and/or adhering cells onto TEM grids manner suitable for while retaining their physiological Here, detailed step-by-step protocol presented on use micropatterning direct promote eukaryotic cell growth grids....