recommended human host cell lines mrc  (ATCC)

Journal: bioRxiv

Article Title: Transcriptomic profiling of human corona virus (HCoV)-229E -infected human cells and genomic mutational analysis of HCoV-229E and SARS-CoV-2

doi: 10.1101/2020.08.17.253682

Figure Lengend Snippet: (A-B) Viral propagation of HCoV-229E. MRC5 cells, 24-hour culture to 80-90% confluency, were infected with 0.01 multiplicity of infection (MOI) of HCoV-229E virus. Cells and virus were grown in Eagle’s Minimum Essential Medium (EMEM) supplemented with 2% fetal calf serum (FCS) in incubated in a humidified incubator with 5% CO2, at 35°C. Total RNA was extracted at the indicated hours post infection (HPI) and viral load was determined in both cells (A) or growth media (B) by qPCR analysis. (C) Active transcripts in HCoV-229E –infected MRC5 cells. Total RNA samples of MRC5-infected cells (from section A) were prepared using the Illumina TruSeq RNA Library Preparation Kit v2 and sequenced in Illumina HiSeq 2500 Sequencer. Software and applications used for sequencing and data analysis are specified in the methods. Total number of upregulated (>2 fold change, red bars) or downregulated (<-2 fold change, blue bars) genes at the indicated HPI was calculated by comparing to gene expression to that measured in uninfected cells.

Article Snippet: HCoV-229E virus was purchased from ATCC (VR740TM) along with its recommended human host cell lines MRC5.

Techniques: Infection, Virus, Incubation, RNA Library Preparation, Software, Sequencing, Gene Expression

Journal: bioRxiv

Article Title: Transcriptomic profiling of human corona virus (HCoV)-229E -infected human cells and genomic mutational analysis of HCoV-229E and SARS-CoV-2

doi: 10.1101/2020.08.17.253682

Figure Lengend Snippet: (A) Canonical pathways enrichment analysis. Analysis was conducted for HCoV-229E-infected MRC5 cells at 6 hours post-infection (HPI) as compared to uninfected cells. Canonical pathways with highest −log p-value (using Benjamini-Hochberg correction) are presented. (B) The number of activated genes was calculated for the eIF2 (I), cell cycle control of chromosomal replication (II), cholesterol biosynthesis (III) and role of BRCA1 in DNA damage response (IV) pathways. Up- and downregulated pathways are presented in red and blue, respectively. (C) Validation of gene expression. Cells were infected with HCoV-229E and total RNA was extracted and treated as described in the Methods section. Relative gene expression was calculated for ORC1 (I), CDK1 (II) mRNA (belong to cell cycle control of chromosomal replication pathway) and ATF3 (III) mRNA (belongs to eIF2 pathway). Fold-change in transcripts levels, as measured by qPCR analysis (Bars, left Y axis), was compared to the results of the NGS analysis (line, right Y axis).

Article Snippet: HCoV-229E virus was purchased from ATCC (VR740TM) along with its recommended human host cell lines MRC5.

Techniques: Infection, Control, Biomarker Discovery, Gene Expression

Journal: bioRxiv

Article Title: Transcriptomic profiling of human corona virus (HCoV)-229E -infected human cells and genomic mutational analysis of HCoV-229E and SARS-CoV-2

doi: 10.1101/2020.08.17.253682

Figure Lengend Snippet: (A) Sum of up and downregulated genes in MRC5 cells at each time point post HCoV-229E infection are presented in red and blue, respectively. (B) Early activated antiviral response genes. Genes that were >2 or <-2 fold change in MRC5 cells at 6 hours post HCoV-229E infection are presented in fold change heat map in following HPI. (C) Validation of gene expression. Cells were infected with HCoV-229E and total RNA was extracted and treated as described in the Methods section. Relative gene expression was calculated for CLU mRNA. Fold-change in transcripts levels, as measured by qPCR analysis (Bars, left Y axis), was compared to the results of the NGS analysis (line, right Y axis).

Article Snippet: HCoV-229E virus was purchased from ATCC (VR740TM) along with its recommended human host cell lines MRC5.

Techniques: Infection, Biomarker Discovery, Gene Expression