HIV-1 reporter pseudovirus expressing SARS-CoV-2 Spike protein and luciferase were generated using two plasmids. pNL4–3. Luc.R-.E- was obtained from the NIH AIDS repository. SARS-CoV-2. SctΔ19 was generated (Geneart) from the full protein sequence of SARS-CoV-2 spike with a deletion of the last 19 amino acids in C-terminal, human-codon optimized and inserted into pcDNA3.4-TOPO [35] (link). Spike plasmid was transfected with X-tremeGENE HP Transfection Reagent (Merck) into HEK-293 T cells, and 24 h post-transfection, cells were transfected with pNL4–3. Luc.R-.E-. Supernatants were harvested 48 h later, filtered with 0.45 μM (Millex Millipore) and stored at −80ºC until use. The p24gag content of all viruses was quantified using an ELISA (Perkin Elmer) and viruses were titrated in HEK-293 T overexpressing the human ACE2.
Sctδ19
Manufactured by Thermo Fisher Scientific
The SctΔ19 is a piece of laboratory equipment designed for scientific research. It is a specialized instrument that serves a core function, though the details of its intended use are not provided in this concise, unbiased description.
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Lab products found in correlation
2 protocols using sctδ19
1
HIV-1 Pseudovirus Expressing SARS-CoV-2 Spike
2
Generating SARS-CoV-2 Spike Pseudoviruses
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HIV-1 reporter pseudoviruses expressing SARS-CoV-2 Spike protein and luciferase were generated using two plasmids. pNL4-3. Luc.R-E-was obtained from the NIH AIDS repository. SARS-CoV-2. SctΔ19 was generated (Geneart) from the full protein sequence of SARS-CoV-2 spike with a deletion of the last 19 amino acids in C-terminal, human-codon optimized and inserted into pcDNA3.4-TOPO (Ou et al., 2020b (link)). Spike plasmid was transfected with X-tremeGENE HP Transfection Reagent (Merck) into HEK-293T cells, and 24 h post-transfection, cells were transfected with pNL4-3. Luc.R-.E-. Supernatants were harvested 48 h later, filtered with 0.45 µM (Millex Millipore) and stored at −80°C until use. Control pseudoviruses were obtained by replacing the spike plasmid by a VSV-G plasmid (kindly provided by Dr Andrea Cimarelli). The p24gag content of all viruses was quantified using an ELISA (Perkin Elmer) and viruses were titrated in HEK-293T overexpressing the human ACE2.
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