A5082022902- Tuberculosis Research and Epidemiology
- Mycobacterium research and diagnosis
- CRISPR and Genetic Engineering
- Antibiotic Resistance in Bacteria
- Pluripotent Stem Cells Research
- Legionella and Acanthamoeba research
- RNA and protein synthesis mechanisms
- Single-cell and spatial transcriptomics
- Bacterial biofilms and quorum sensing
- Congenital heart defects research
- Vibrio bacteria research studies
- Toxoplasma gondii Research Studies
- Genomic variations and chromosomal abnormalities
- Diagnosis and treatment of tuberculosis
- Amyotrophic Lateral Sclerosis Research
- Venomous Animal Envenomation and Studies
- Advanced biosensing and bioanalysis techniques
- Heme Oxygenase-1 and Carbon Monoxide
- Neuroinflammation and Neurodegeneration Mechanisms
- Parkinson's Disease Mechanisms and Treatments
- Cytomegalovirus and herpesvirus research
- Genomics and Rare Diseases
- Oral and gingival health research
- Transplantation: Methods and Outcomes
- Parvovirus B19 Infection Studies
Jackson Laboratory
2018-2025
Yale University
2002-2023
KU Leuven
2015
University of North Carolina at Chapel Hill
2005-2010
Duke University
2010
Duke University Hospital
2010
Duke Medical Center
2010
Indiana University School of Medicine
2008-2009
University of Wisconsin–Madison
2008
James Madison University
2003
Human induced pluripotent stem cell (iPSC) lines are a powerful tool for studying development and disease, but the considerable phenotypic variation between makes it challenging to replicate key findings integrate data across research groups. To address this issue, we sub-cloned candidate human iPSC deeply characterized their genetic properties using whole genome sequencing, genomic stability upon CRISPR-Cas9-based gene editing, including differentiation commonly used types. These studies...
Coxiella burnetii is an intracellular pathogen that replicates in a lysosome-derived vacuole. The molecular mechanisms used by this bacterium to create pathogen-occupied vacuole remain largely unknown. Here, we conducted visual screen on arrayed library of C. NMII transposon insertion mutants identify genes required for biogenesis mature Coxiella-containing (CCV). Mutants defective Dot/Icm secretion system function or the PmrAB regulatory were incapable replication. Several with growth...
The human pathogen Coxiella burnetii encodes a type IV secretion system called Dot/Icm that is essential for intracellular replication. delivers bacterial effector proteins into the host cytosol during infection. delivered by C. are predicted to have important functions infection, but when these needed infection has not been clearly defined. Here, we use reporter consisting of fusion β-lactamase enzyme (BlaM) fused study protein translocation system. Translocation BlaM CBU0077, CBU1823 and...
ABSTRACT Coxiella burnetii is an intracellular pathogen that replicates within a lysosome-like vacuole. A Dot/Icm type IVB secretion system used by C. to translocate effector proteins into the host cytosol likely modulate factor function. To identify determinants required for growth, genome-wide screen was performed using gene silencing small interfering RNA (siRNA). Replication of measured immunofluorescence microscopy in siRNA-transfected HeLa cells. Newly identified factors included...
ABSTRACT The twin-arginine translocation (Tat) pathway exports folded proteins across the bacterial cytoplasmic membrane and is responsible for proper extracytoplasmic localization of involved in a variety cellular functions, including pathogenesis. Mycobacterium tuberculosis smegmatis genomes contain open reading frames with homology to components Tat export system (TatABC) as well potential Tat-exported possessing N-terminal signal sequences characteristic motif. Due importance exported...
Abstract Microglia play a pivotal role in neurodegenerative disease pathogenesis, but the mechanisms underlying microglia dysfunction and toxicity remain to be elucidated. To investigate effect of disease-linked genes on intrinsic properties microglia, we studied microglia-like cells derived from human induced pluripotent stem (iPSCs), termed iMGs, harboring mutations profilin-1 (PFN1) that are causative for amyotrophic lateral sclerosis (ALS). ALS-PFN1 iMGs exhibited evidence lipid...
ABSTRACT The use of antibiotic resistance analysis (ARA) for microbial source tracking requires the generation a library isolates collected from known sources in watershed. size and composition are critical determining if it represents diversity patterns found This study was performed to determine that an ARA needs be representative watersheds which will used libraries different can merged create multiwatershed libraries. Fecal samples human, domesticated, wild animal were six Virginia...
The twin-arginine translocation (Tat) pathway is a system used by some bacteria to export proteins out from the cytosol cell surface or extracellular environment. A functional Tat exists in important human pathogen Mycobacterium tuberculosis. Identification of substrates exported can help define role that this plays physiology and pathogenesis M. Here we reporter export, truncated beta-lactamase, 'BlaC, experimentally identify tuberculosis with signal sequences. Of 13 identified, one lacks...
Anaplasma phagocytophilum is an obligate intracellular bacterium that infects neutrophils to reside within a host cell-derived vacuole. The A. phagocytophilum-occupied vacuole (ApV) fails mature along the endocytic pathway and non-fusogenic with lysosomes. Rab GTPases regulate membrane traffic. To better understand how modulates ApV's selective fusogencity, we examined localization of 20 green fluorescent protein (GFP) or red (RFP)-tagged in phagocytophilum-infected HL-60 cells. GFP-Rab4A,...
In bacteria, the majority of exported proteins are transported by general Sec pathway from their site synthesis in cytoplasm across cytoplasmic membrane. The essential SecA ATPase powers this Sec-mediated export. Mycobacteria possess two nonredundant homologs: SecA1 and SecA2. pathogenic Mycobacterium tuberculosis nonpathogenic model mycobacterium smegmatis, is for protein export "housekeeping" SecA, whereas SecA2 an accessory that exports a specific subset proteins. M. plays role virulence....
The ‘PTC+1’ knockout strategy offers a robust and efficient approach to eliminate gene function in human induced pluripotent stem cells (iPSCs). method utilizes precise genome engineering introduce premature termination codon (PTC) degenerate base insertion (+1) within an early coding exon that is shared across all transcript isoforms. Here, we describe the protocol for generation of alleles KOLF2.2J iPSCs achieve targeted knockout. Using nucleofection, co-deliver synthetic oligonucleotide...
Precise deletions are performed in human induced pluripotent stem cells (iPSC) by Cas9 RNP-mediated homology directed repair with oligonucleotide templates that span the desired breakpoints. We aimed to delete 1) a ‘critical exon’ (CE) common all transcript isoforms that, when deleted, causes frameshift coding sequence or 2) up 100 kb of containing most protein (whole-gene knockout, KO). A bridging oligo two degenerate nucleotides at breakpoint was designed promote HDR and barcoding precise...
The specification of extra-embryonic mesenchyme (ExMC) is a prime example developmental divergence between mouse and human. Derived from definitive mesoderm during gastrulation, the human ExMC first appears at peri-implantation prior to gastrulation therefore its cellular origin, still unknown, must differ. In pluripotent stem cell model, we report that shares progenitor cells with trophoblast, suggesting trophectoderm origin. This ability form extend trophoblast lines. We define HAND1 as an...
ABSTRACT The exported proteins of Mycobacterium tuberculosis that are localized at the bacterial cell surface or secreted into environment ideally situated to interact with host factors and function in virulence. In this study, we constructed a novel β-lactamase reporter transposon used it directly M. for genome-wide identification proteins. From 177 β-lactam-resistant mutants, identified 111 different majority these have no known function, nearly half proteins, our demonstration they when...
Mycobacterium tuberculosis is an intracellular pathogen that able to avoid destruction by host immune defences. Exported proteins of M. tuberculosis, which include localized the bacterial surface or secreted into extracellular environment, are ideally situated interact with factors. As a result, these attractive candidates for virulence factors, drug targets and vaccine components. Here we describe β-lactamase reporter system capable identifying exported during growth in cells. Because...
Abstract Human induced pluripotent stem cell (iPSC) lines are a powerful tool for studying development and disease, but the considerable phenotypic variation between makes it challenging to replicate key findings integrate data across research groups. To address this issue, we sub-cloned candidate iPSC deeply characterised their genetic properties using whole genome sequencing, genomic stability upon CRISPR/Cas9-based gene editing, including differentiation commonly-used types. These studies...