hcov 229e  (ATCC)

Journal: iScience

Article Title: Broad-spectrum antiviral activity of antisense oligonucleotides targeting GBF1 against SARS-CoV-2 and influenza viruses

doi: 10.1016/j.isci.2026.114851

Figure Lengend Snippet: Identification of host factors involved in SARS-CoV-2 and HCoV-229E replication (A) Schematic overview of the siRNA screening. HEK293 A/T cells were seeded in 24-well plates and transfected twice with siRNAs targeting 91 host genes previously identified as being involved in influenza virus replication. Cells were then infected with SARS-CoV-2 (100 plaque-forming unit [PFU]/100 μL) one day after the second transfection. Supernatants were collected at 2 days post-infection (dpi) and titrated by plaque assay. (B) Results of the SARS-CoV-2 siRNA screen. The 91 host factors were divided into four batches, each including a non-targeting siRNA as a negative control (N) and siRNA targeting SARS-CoV-2 nsp12 as a positive control (P). Viral titers were calculated based on the difference between each siRNA and its corresponding negative control. Each dot represents the mean of duplicate wells from a single independent experiment. Data are presented as the mean ± standard deviation (SD) of at least three independent experiments. (C) The seven host factors identified in the SARS-CoV-2 siRNA screen were further examined in HCoV-229E, along with the negative control siRNA (N) and positive control siRNA (P) targeting the HCoV-229E N gene. Following the same method described in (A), MRC-5 cells were infected with HCoV-229E (50 tissue culture infectious dose (TCID 50 )/100 μL). Supernatants were collected at 3 dpi and titrated by TCID 50 assay. Viral titers were calculated based on the difference between each siRNA and the negative control. Each dot represents the mean of duplicate wells from a single independent experiment. Data are presented as the mean ± SD of three independent experiments. (D) Cell viability was measured in duplicate wells across two independent experiments using CellTiter-Glo following siRNA transfection. See also and .

Article Snippet: HCoV-229E , ATCC , VR-740.

Techniques: Transfection, Virus, Infection, Plaque Assay, Negative Control, Positive Control, Standard Deviation

Journal: Analytical chemistry

Article Title: Viral Membrane-Targeting Amphipathic Helical Peptide-Based Fluorogenic Probes for the Analysis of Infectious Titers of Enveloped Viruses.

doi: 10.1021/acs.analchem.4c04852

Figure Lengend Snippet: Figure 2. (A) Sequences of AH peptides used for the screening of the binders toward the HCoV-229E virus. X denotes norleucine used instead of methionine in the wild sequence (Table S1). (B) Chemical structure of M2-NR. (C) Fluorescence response of the AH peptide conjugated with the NR unit (2.0 μM) for 1.7 × 109 particles/mL HCoV-229E virus. Measurements were performed in PBS buffer at 25 °C with excitation at 552 nm. Analysis, 640 nm.

Article Snippet: The HCoV strain 229E was obtained as shown in the literature or purchased from ATCC (VR-740).

Techniques: Virus, Sequencing, Fluorescence

Journal: Analytical chemistry

Article Title: Viral Membrane-Targeting Amphipathic Helical Peptide-Based Fluorogenic Probes for the Analysis of Infectious Titers of Enveloped Viruses.

doi: 10.1021/acs.analchem.4c04852

Figure Lengend Snippet: Figure 3. (A) Fluorescence spectra of M2-NR (2.0 μM) in the absence and presence of 1.7 × 109 particles/mL HCoV-229E. The response to 1.7 × 109 particles/mL AAV-2 is also shown. (B) Effect of protease K (0.1 mg/mL) or Triton-X (0.1%) treatment on the fluorescence response of M2- NR (2.0 μM) to 1.7 × 109 particles/mL HCoV-229E. (C) Fluorescence response of M2-NR (2.0 μM) to serially diluted HCoV-229E (0−1.7 × 109 particles/mL). Excitation, 552 nm. Analysis, 640 nm. Temperature, 25 °C.

Article Snippet: The HCoV strain 229E was obtained as shown in the literature or purchased from ATCC (VR-740).

Techniques: Fluorescence

Journal: Analytical chemistry

Article Title: Viral Membrane-Targeting Amphipathic Helical Peptide-Based Fluorogenic Probes for the Analysis of Infectious Titers of Enveloped Viruses.

doi: 10.1021/acs.analchem.4c04852

Figure Lengend Snippet: Figure 4. (A) Linear relationship between the HCoV-229E particle concentration (particles/mL) by the M2-NR response and the virus titer based on the TCID50 assay. (B) Comparison of the virus titer calculated by an M2-NR fluorescence assay and those determined by TCID50 assay; [M2-NR] = 2.0 μM.

Article Snippet: The HCoV strain 229E was obtained as shown in the literature or purchased from ATCC (VR-740).

Techniques: Concentration Assay, Virus, TCID50 Assay, Comparison, Fluorescence

Journal: Analytical chemistry

Article Title: Viral Membrane-Targeting Amphipathic Helical Peptide-Based Fluorogenic Probes for the Analysis of Infectious Titers of Enveloped Viruses.

doi: 10.1021/acs.analchem.4c04852

Figure Lengend Snippet: Figure 5. (A) Fluorescence response of M2-NR to serially diluted virus (HCoV-229EHuh‑7, IAV, HSV-1, or VSV-G LV). (B) Comparison of the virus titer (HCoV-229EHuh‑7 (TCID50/mL), IAV (TCID50/mL), HSV (TCID50/mL), or VSV-G LV (IFU/mL)), calculated by the M2-NR fluorescence assay and those determined by cell-based assays; [M2-NR] = 2.0 μM.

Article Snippet: The HCoV strain 229E was obtained as shown in the literature or purchased from ATCC (VR-740).

Techniques: Fluorescence, Virus, Comparison