A5032772458- Brucella: diagnosis, epidemiology, treatment
- RNA Interference and Gene Delivery
- Bacteriophages and microbial interactions
- Escherichia coli research studies
- CRISPR and Genetic Engineering
- Advanced biosensing and bioanalysis techniques
- Salmonella and Campylobacter epidemiology
- Molecular Biology Techniques and Applications
- Cellular Mechanics and Interactions
- Monoclonal and Polyclonal Antibodies Research
- Bacterial biofilms and quorum sensing
- Mesenchymal stem cell research
- Integrated Circuits and Semiconductor Failure Analysis
- Polyomavirus and related diseases
- Cardiomyopathy and Myosin Studies
- RNA Research and Splicing
- Yersinia bacterium, plague, ectoparasites research
- Antimicrobial agents and applications
- Bacterial Genetics and Biotechnology
- Microtubule and mitosis dynamics
- Advanced Fluorescence Microscopy Techniques
- Tardigrade Biology and Ecology
- Hematopoietic Stem Cell Transplantation
- Galectins and Cancer Biology
- Wound Healing and Treatments
Nanyang Technological University
2021
University of Basel
2010-2019
Temasek Life Sciences Laboratory
2010-2012
Significance Pathogens can enter into human cells using a variety of specific mechanisms, often hitchhiking on naturally existing transport pathways. To uncover parts the host machinery that are required for entry, scientists conduct infection screens in cultured cells. In these screens, genes systematically inactivated by short RNA oligos, designed to bind and inactivate mRNA molecules. Here, we show many oligos additionally unintended targets as well, this effect overall dominates...
ABSTRACT Entry of the facultative intracellular pathogen Brucella into host cells results in formation endosomal Brucella-containing vacuoles (eBCVs) that initially traffic along endocytic pathway. eBCV acidification triggers expression a type IV secretion system translocates bacterial effector proteins cells. This interferes with lysosomal fusion eBCVs and supports their maturation to replicative (rBCVs). Bacteria replicate rBCVs large numbers, eventually occupying most cytoplasmic volume....
Large-scale RNAi screening has become an important technology for identifying genes involved in biological processes of interest. However, the quality large-scale is often deteriorated by off-targets effects. In order to find statistically significant effector pathogen entry, we systematically analyzed entry pathways human host cells eight pathogens using image-based kinome-wide siRNA screens with siRNAs from three vendors. We propose a Parallel Mixed Model (PMM) approach that simultaneously...
Small interfering RNAs (siRNAs) exhibit strong off-target effects, which confound the gene-level interpretation of RNA interference screens and thus limit their utility for functional genomics studies. Here, we present gespeR, a statistical model reconstructing individual, gene-specific phenotypes. Using 115,878 siRNAs, single pooled, from three companies in pathogen infection screens, demonstrate that deconvolution image-based phenotypes substantially improves reproducibility between...
Methods enabling the delivery of proteins into eukaryotic cells are essential to address protein functions. Here we propose broad applications cell biology for a tool based on bacterial type III secretion (T3S). We show that bacterial, viral, and human proteins, fused N-terminal fragment Yersinia enterocolitica T3S substrate YopE, effectively delivered target in fast controllable manner via injectisome extracellular bacteria. This method enables functional interaction studies by simultaneous...
With >300,000 new cases of human brucellosis annually, Brucella is regarded as one the most important zoonotic bacterial pathogens worldwide. The agent causing resides inside host cells within vacuoles termed Brucella- containing (BCVs). Although a few components required to escape degradative lysosomal pathway and establish ER-derived replicative BCV (rBCV) have already been identified, global understanding this highly coordinated process still partial, many factors remain unknown. To...
The actin cross-linking protein, α-actinin, plays a crucial role in mediating furrow ingression during cytokinesis. However, the mechanism by which its dynamics are regulated this process is poorly understood. Here we have investigated of calcium sensitivity α-actinin regulation generating functional calcium-insensitive mutant (EFM). GFP-tagged EFM (EFM-GFP) localized to equatorial regions cell division. maximal accumulation EFM-GFP was significantly smaller comparison α-actinin-GFP when it...
Perturbation of gene expression by means synthetic small interfering RNAs (siRNAs) is a powerful way to uncover function. However, siRNA technology suffers from sequence-specific off-target effects and limitations in knock-down efficiency. In this study, we assess further problem: unintended transfections on cellular fitness/proliferation. We show that the nucleotide compositions siRNAs at specific positions have reproducible growth-restricting mammalian cells culture. This likely distinct...
Brucella species are facultative intracellular pathogens that infect animals as their natural hosts. Transmission to humans is most commonly caused by direct contact with infected or ingestion of contaminated food and can lead severe chronic infections. invade professional non-professional phagocytic cells replicates within endoplasmic reticulum (ER)-derived vacuoles. The host factors required for entry into cells, avoidance lysosomal degradation, replication in the ER-like compartment...
ABSTRACT Staphylococcus aureus (SA) is the most common bacterial species in chronic wounds. However, there a lack of understanding how SA secretions affect cell biology during healing process. We studied effects biofilm-secretions from strain SA29213 on 3T3 fibroblasts. wound isolate and widely used as reference strain. series concentrations biofilm-conditioned media (BCM) found 100% BCM lethal within 10 h. Cells survived ≤75% but rate closure scratch assays was reduced. Treatment with 75%...
Brucella species are facultative intracellular pathogens that infect animals as their natural hosts. Transmission to humans is most commonly caused by direct contact with infected or ingestion of contaminated food and can lead severe chronic infections. invade professional non-professional phagocytic cells replicates within endoplasmic reticulum (ER)-derived vacuoles. The host factors required for entry into cells, avoidance lysosomal degradation, replication in the ER-like compartment...
Abstract Brucella, the causing agent of brucellosis, is a major zoonotic pathogen with worldwide distribution. Brucella resides and replicates inside infected host cells in membrane-bound compartments called BCVs ( Brucella- containing vacuoles). Following uptake, eBCVs (endosomal BCVs) that gradually mature from early to late endosomal features. Through poorly understood process key intracellular lifestyle eBCV escapes fusion lysosomes by transitioning rBCV (replicative BCV), replicative...