Other Publications (1)
Articles by Andre Dharmawan in JoVE
Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications Cameron A. Smurthwaite1, Wesley Williams1, Alexandra Fetsko1, Darin Abbadessa1, Zachary D. Stolp1, Connor W. Reed1, Andre Dharmawan1, Roland Wolkowicz1 1Department of Biology, San Diego State University Since the discovery of the green fluorescent protein gene, fluorescent proteins have impacted molecular cell biology. This protocol describes how expression of distinct fluorescent proteins through genetic engineering is used for barcoding individual cells. The procedure enables tracking distinct populations in a cell mixture, which is ideal for multiplexed applications.
Other articles by Andre Dharmawan on PubMed
A Multiplexed Cell-Based Assay for the Identification of Modulators of Pre-Membrane Processing As a Target Against Dengue Virus Journal of Biomolecular Screening. Feb, 2015 | Pubmed ID: 25724189 The DenV pre-membrane protein (prM) is a crucial chaperone for the viral envelope protein, preventing premature fusion with vesicles during viral export. prM molecules in immature particles are cleaved by host proteases, leading to mature fusogenic virions. Blockade of prM cleavage would restrict fusion and represents a novel druggable opportunity against DenV. We have thus established a cell-based platform to monitor prM processing that relies on an engineered two-tag scaffold that travels to the cell surface through the secretory pathway. The assay discriminates between a single cell-surface tag when prM is cleaved and two tags when it is not, as detected through fluorescent-coupled antibodies by flow cytometry. The assay, miniaturized into a 96-well plate format, was multiplexed with the HIV-1 envelope boundary, also cleaved in the same pathway. A pilot screen against 1280 compounds was executed, leading to the identification of a potential active and corroborating the robustness of our assay for large-scale screening. We describe for the first time a cell-based assay that monitors DenV prM processing within the classical secretory pathway, which was exploited to identify a potential novel drug against DenV.