Editing SARS2 Pseudovirus (section)

== Pseudotyping lentiviral particles with SARS-CoV-2 Spike protein for neutralization assays ==



[[File:Pseudovirus.png]]



Plasmids

pHAGE2-EF1aInt-ACE2-WT (BEI catalog number NR52512): lentiviral backbone plasmid expressing&nbsp;the human ACE2 gene (GenBank ID for human ACE2 is NM_021804) under an EF1a promoter with an&nbsp;intron to increase expression.<br/>• HDM-IDTSpike-fixK-HA-tail (BEI catalog number NR52513): plasmid expressing under a CMV&nbsp;promoter the Spike from SARS-CoV-2 strain Wuhan-Hu-1 (Genbank NC_045512) codon-optimized&nbsp;using IDT, with the Spike cytoplasmic tail replaced by that from the HA protein of A/WSN/1933&nbsp;(H1N1) influenza, and the Kozak sequence in the plasmid fixed compared to an earlier version of this&nbsp;plasmid.<br/>• HDM-IDTSpike-fixK (BEI catalog number NR-52514): plasmid expressing under a CMV promoter the&nbsp;Spike from SARS-CoV-2 strain Wuhan-Hu-1 (Genbank NC_045512) codon-optimized using IDT and&nbsp;the Kozak sequence in the plasmid fixed compared to an earlier version of this plasmid.<br/>• HDM-nCoV-Spike-IDTopt-ALAYT (BEI catalog number NR-52515): plasmid expressing under a CMV&nbsp;promoter the Spike from SARS-CoV-2 strain Wuhan-Hu-1 (Genbank NC_045512) codon-optimized&nbsp;using IDT, with the Spike containing two mutations in the cytoplasmic tail such that the last five amino<br/>acids are ALAYT.<br/>• pHAGE-CMV-Luc2-IRES-ZsGreen-W (BEI catalog number NR-52516): lentiviral backbone plasmid&nbsp;that uses a CMV promoter to express luciferase followed by an IRES and ZsGreen.<br/>• HDM-Hgpm2 (BEI catalog number NR-52517): lentiviral helper plasmid expressing HIV Gag-Pol&nbsp;under a CMV promoter.<br/>• HDM-tat1b (NR-52518): lentiviral helper plasmid expressing HIV Tat under a CMV promoter.<br/>• pRC-CMV-Rev1b (NR-52519): lentiviral helper plasmid expressing HIV Rev under a CMV promoter.<br/>• pHAGE2-CMV-ZsGreen-W (NR-52520): lentiviral backbone plasmid that uses a CMV promoter to express ZsGreen.