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Genome-wide knockout screen of OC43/SARS-CoV2-infected HEK293-hACE2 and Vero E6 cells

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Homo sapiens Chlorocebus sabaeus

PROVIDER: GSE177545 | GEO | 2021/12/22

REPOSITORIES: GEO

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Genome-wide knockout screen of OC43/SARS-CoV2-infected Vero E6 cells

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Genome-wide knockout screen of OC43/SARS-CoV2-infected HEK293-hACE2 cells II

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Genome-wide knockout screen of OC43/SARS-CoV2-infected HEK293-hACE2 and Vero E6 cells

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Genome-wide knockout screen of OC43/SARS-CoV2-infected Vero E6 cells

Project description:Genome-wide knockout screen of OC43/SARS-CoV2-infected Vero E6 cells

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Genome-wide knockout screen of OC43/SARS-CoV2-infected HEK293-hACE2 cells I

Project description:Genome-wide knockout screen of OC43/SARS-CoV2-infected HEK293-hACE2 cells I

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Genome-wide knockout screen of OC43/SARS-CoV2-infected HEK293-hACE2 cells II

Project description:Genome-wide knockout screen of OC43/SARS-CoV2-infected HEK293-hACE2 cells II

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Gene expression of SARS-CoV-2-infected Vero E6 cells

Project description:We conducted a high-throughput drug repositioning screen using the LOPAC®1280 and the ReFRAME drug libraries to identify existing drugs that harbor antiviral activity against SARS-CoV-2, in a Vero E6 cell-based assay. We additionally performed RNA sequencing on control and SARS-CoV-2 infected Vero E6 cells to study the biological changes after SARS-CoV-2 infection and to elucidate the potential mechanisms underlying the positive hits identified from our high-throughput screen. Vero E6 cells were either mock-infected or infected with SARS-CoV-2 USA-WA1/2020 (MOI = 0.3) with three replicates. Cells were harvested 24 hours after infection, and total RNA was extracted using the Qiagen® RNeasy® Plus Mini Kit. The quality of the extracted RNA was assessed with the Agilent® 2100 Bioanalyzer. Libraries were prepared from total RNA following ribosome RNA depletion using standard protocol according to Illumina®. Total RNA sequencing was then performed on the Illumina® NextSeq system; 150bp paired-end runs were performed and 100 million raw reads per sample were generated.

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Imaging of SARS-CoV-2 infected Vero E6 cells by helium ion microscopy.

Project description:Helium ion microscopy (HIM) offers the opportunity to obtain direct views of biological samples such as cellular structures, virus particles, and microbial interactions. Imaging with the HIM combines sub-nanometer resolution, large depth of field, and high surface sensitivity. Due to its charge compensation capability, the HIM can image insulating biological samples without additional conductive coatings. Here, we present an exploratory HIM study of SARS-CoV-2 infected Vero E6 cells, in which several areas of interaction between cells and virus particles, as well as among virus particles, were imaged. The HIM pictures show the three-dimensional appearance of SARS-CoV-2 and the surface of Vero E6 cells at a multiplicity of infection of approximately 1 with great morphological detail. The absence of a conductive coating allows for a distinction between virus particles bound to the cell membrane and virus particles lying on top of the membrane. After prolonged imaging, it was found that ion-induced deposition of hydrocarbons from the vacuum renders the sample sufficiently conductive to allow for imaging even without charge compensation. The presented images demonstrate the potential of the HIM in bioimaging, especially for the imaging of interactions between viruses and their host organisms.

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