Search bioRxiv⌕ Search

Biology subjects

Hasson, S.

Publications and source records attributed to Hasson, S..

3 recordsLinked to original sources

Identification of bacteria by poly-aromatic hydrocarbons biosensors

Human health is consistently threatened by different species of pathogenic bacteria. To fight the spread of diseases, it is important to develop rapid methods for bacterial identification. Over the years, different kinds of biosensors were developed for this cause. Another environmental risk are poly-aromatic hydrocarbons (PAHs) that may be emitted from industrial facilities and pollute environmental water and soil. One of the methods for their purification is conducted by the addition of bacteria that can degrade the PAHs, while the bacteria itself can be filtrated at the end of the process. Although many studies reported monitoring of the PAHs degradation by fluorescence, not much attention was dedicated to studying the influence of the PAHs on the intrinsic fluorescence of the degrading bacteria. In this work, we apply synchronous fluorescence (SF) measurements to study the ability of the 5 PAHs: 9-Antracene carboxylic acid (9ACA), Pyrene, Perylene, Pentacene, and Chrysene to interact with bacteria and change its fluorescence spectra. We show that upon incubation of each PAH with the bacterium E.coli only the 2 PAHs 9ACA and Perylene cause an intensity decrease in the emission at {lambda} = 300 - 375 nm, which derives from the emission of Tyrosine and Tryptophane (TT). Also, we show that upon incubation of 9ACA and Perylene with 5 different pathogenic bacteria, the intensity increase or decrease in the TT emission is unique to each bacterial species. Based on this observation, we suggest that the PAHs 9ACA and Perylene can be utilized as biosensors for bacterial identification.

microbiology↗

A Novel Tmem119-tdTomato Reporter Mouse Model for Studying Microglia in the Central Nervous System

Microglia are resident immune cells of the central nervous system (CNS). The exact role of microglia in the physiopathology of CNS disorders is not clear due to lack of tools to discriminate between CNS resident and infiltrated innate immune cells. Here, we present a novel reporter mouse model targeting a microglia-specific marker (TMEM119) for studying the function of microglia in health and disease. By placing a reporter cassette (GSG-3xFlag-P2A-tdTomato) between the coding sequence of exon 2 and 3UTR of the Tmem119 gene using CRISPR/Cas9 technology, we generated a Tmem119-tdTomato knock-in mouse strain. Gene expression assay showed no difference of endogenous Tmem119 mRNA level in the CNS of Tmem119tdTomato/+ relative to control Wild-type mice. The cells expressing tdTomato-were recognized by immunofluorescence staining using commercially available anti-TMEM119 antibodies. Using immunofluorescence and flow cytometry techniques, tdTomato+ cells were detected throughout the CNS, but not in peripheral tissues of adult Tmem119tdTomato/+ mice. In addition, aging does not seem to influence TMEM119 expression as tdTomato+ cells were detectable in the CNS of older mice (300 and 540 days old). Further immunofluorescence characterization shows that the tdTomato+ cells were highly colocalized with Iba1+ cells (microglia and macrophages) in the brain, but not with NeuN- (neurons), GFAP- (astrocytes) or Olig2- (oligodendrocytes) labeled cells. Moreover, flow cytometry analysis of brain tissues of adult mice demonstrates that the majority of microglial CD45lowCD11b+ cells (96.6%) are tdTomato positive. Functionally, using a laser-induced injury model, we measured time-lapse activation of tdTomato-labeled microglia by transcranial two-photon microscopy in live Tmem119tdTomato/+ mice. Taken together, the Tmem119-tdTomato reporter mouse model will serve as a valuable tool to specifically study the role of microglia in health and disease.

immunology↗

A systematic survey of human tissue-specific gene expression and splicing reveals new opportunities for therapeutic target identification and evaluation

Differences in the expression of genes and their splice isoforms across human tissues are fundamental factors to consider for therapeutic target evaluation. To this end, we conducted a transcriptome-wide survey of tissue-specific gene expression and splicing events in the unprecedented collection of 8527 high-quality RNA-seq samples from the GTEx project, covering 36 human peripheral tissues and 13 brain subregions. We derived a weighted tissue-specificity scoring scheme accounting for the similarity of related tissues and inherent variability across individual samples. We showed that ~50.6% of all annotated human genes show tissue-specific expression, including many low abundance transcripts vastly underestimated by previous array-based expression atlases. As utilities for drug discovery, we demonstrated that tissue-specificity is a highly desirable attribute of validated drug targets and tissue-specificity can be used to prioritize disease-associated genes from genome-wide association studies (GWAS). Using brain striatum-specific gene expression as an example, we provided a template to leverage tissue-specific gene expression to identify novel therapeutic targets. Mining of tissue-specific splicing further reveals new opportunities for tissue-specific targeting. Thus, the high quality transcriptome atlas provided by the GTEx is an invaluable resource for drug discovery and systematic analysis anchored on the human tissue specific gene expression provides a promising avenue to identify novel therapeutic target hypotheses.

genomics↗