Retroviral:Article Title: A prenylated dsRNA sensor protects against severe COVID-19.
Article Snippet: .. Retroviral vectors and cell modification The lentiviral vector pSCRPSY (KT368137.1) has been previously described (5). pLV-EF1aIRES-Puro (Addgene plasmid #85132) or pLVEF1a-IRES-Blast (Addgene plasmid #85133) were modified by polymerase chain reaction (PCR) amplifying the TagRFP ORF (using pSCRPSY as template) flanked by directional SfiI sites, which were further flanked by BamHI and EcoRI restriction sites (forward oligo: 5′-CTC TCG GAT CCG GCC GAG AGG GCC ATG AGC GAG CTG ATT AAG-3′ and reverse oligo: 5′-CTC TCG AAT TCG GCC AGA GAG GCC TCA CTT GTG CCC CAG-3′), and the BamHI-EcoRI fragment was subcloned into the vectors to create the modified pLVEF1a-IRES-Puro-SfiI-TagRFP or pLV-EF1aIRES-Blast-SfiI-TagRFP constructs. .. The cDNA corresponding to the ORFs of the following OAS genes (GenBank accession number): OAS1p46 (NM_016816), human OAS3 (NM_006187), mouse Oas1A (NM_145211), bovine OAS1X (NM_178108), bovine OAS1Y (NM_001040606), bovine OAS1Z (AY650038), P. kuhlii (XM_036409709.1), C. dromedarius (XM_031443284), R. ferrumequinum OAS1 (short isoform: XM_033097132 / long isoform: ENSRFET00010016745), and P. alecto (NM_001290162) were synthesized as gene blocks with flanking SfiI sites (IDT DNA), and the SfiI fragment was subcloned into the modified pLV-EF1a-IRES-Puro-SfiI plasmid.
Modification:Article Title: A prenylated dsRNA sensor protects against severe COVID-19.
Article Snippet: .. Retroviral vectors and cell modification The lentiviral vector pSCRPSY (KT368137.1) has been previously described (5). pLV-EF1aIRES-Puro (Addgene plasmid #85132) or pLVEF1a-IRES-Blast (Addgene plasmid #85133) were modified by polymerase chain reaction (PCR) amplifying the TagRFP ORF (using pSCRPSY as template) flanked by directional SfiI sites, which were further flanked by BamHI and EcoRI restriction sites (forward oligo: 5′-CTC TCG GAT CCG GCC GAG AGG GCC ATG AGC GAG CTG ATT AAG-3′ and reverse oligo: 5′-CTC TCG AAT TCG GCC AGA GAG GCC TCA CTT GTG CCC CAG-3′), and the BamHI-EcoRI fragment was subcloned into the vectors to create the modified pLVEF1a-IRES-Puro-SfiI-TagRFP or pLV-EF1aIRES-Blast-SfiI-TagRFP constructs. .. The cDNA corresponding to the ORFs of the following OAS genes (GenBank accession number): OAS1p46 (NM_016816), human OAS3 (NM_006187), mouse Oas1A (NM_145211), bovine OAS1X (NM_178108), bovine OAS1Y (NM_001040606), bovine OAS1Z (AY650038), P. kuhlii (XM_036409709.1), C. dromedarius (XM_031443284), R. ferrumequinum OAS1 (short isoform: XM_033097132 / long isoform: ENSRFET00010016745), and P. alecto (NM_001290162) were synthesized as gene blocks with flanking SfiI sites (IDT DNA), and the SfiI fragment was subcloned into the modified pLV-EF1a-IRES-Puro-SfiI plasmid.
Plasmid Preparation:Article Title: A prenylated dsRNA sensor protects against severe COVID-19.
Article Snippet: .. Retroviral vectors and cell modification The lentiviral vector pSCRPSY (KT368137.1) has been previously described (5). pLV-EF1aIRES-Puro (Addgene plasmid #85132) or pLVEF1a-IRES-Blast (Addgene plasmid #85133) were modified by polymerase chain reaction (PCR) amplifying the TagRFP ORF (using pSCRPSY as template) flanked by directional SfiI sites, which were further flanked by BamHI and EcoRI restriction sites (forward oligo: 5′-CTC TCG GAT CCG GCC GAG AGG GCC ATG AGC GAG CTG ATT AAG-3′ and reverse oligo: 5′-CTC TCG AAT TCG GCC AGA GAG GCC TCA CTT GTG CCC CAG-3′), and the BamHI-EcoRI fragment was subcloned into the vectors to create the modified pLVEF1a-IRES-Puro-SfiI-TagRFP or pLV-EF1aIRES-Blast-SfiI-TagRFP constructs. .. The cDNA corresponding to the ORFs of the following OAS genes (GenBank accession number): OAS1p46 (NM_016816), human OAS3 (NM_006187), mouse Oas1A (NM_145211), bovine OAS1X (NM_178108), bovine OAS1Y (NM_001040606), bovine OAS1Z (AY650038), P. kuhlii (XM_036409709.1), C. dromedarius (XM_031443284), R. ferrumequinum OAS1 (short isoform: XM_033097132 / long isoform: ENSRFET00010016745), and P. alecto (NM_001290162) were synthesized as gene blocks with flanking SfiI sites (IDT DNA), and the SfiI fragment was subcloned into the modified pLV-EF1a-IRES-Puro-SfiI plasmid.
Polymerase Chain Reaction:Article Title: A prenylated dsRNA sensor protects against severe COVID-19.
Article Snippet: .. Retroviral vectors and cell modification The lentiviral vector pSCRPSY (KT368137.1) has been previously described (5). pLV-EF1aIRES-Puro (Addgene plasmid #85132) or pLVEF1a-IRES-Blast (Addgene plasmid #85133) were modified by polymerase chain reaction (PCR) amplifying the TagRFP ORF (using pSCRPSY as template) flanked by directional SfiI sites, which were further flanked by BamHI and EcoRI restriction sites (forward oligo: 5′-CTC TCG GAT CCG GCC GAG AGG GCC ATG AGC GAG CTG ATT AAG-3′ and reverse oligo: 5′-CTC TCG AAT TCG GCC AGA GAG GCC TCA CTT GTG CCC CAG-3′), and the BamHI-EcoRI fragment was subcloned into the vectors to create the modified pLVEF1a-IRES-Puro-SfiI-TagRFP or pLV-EF1aIRES-Blast-SfiI-TagRFP constructs. .. The cDNA corresponding to the ORFs of the following OAS genes (GenBank accession number): OAS1p46 (NM_016816), human OAS3 (NM_006187), mouse Oas1A (NM_145211), bovine OAS1X (NM_178108), bovine OAS1Y (NM_001040606), bovine OAS1Z (AY650038), P. kuhlii (XM_036409709.1), C. dromedarius (XM_031443284), R. ferrumequinum OAS1 (short isoform: XM_033097132 / long isoform: ENSRFET00010016745), and P. alecto (NM_001290162) were synthesized as gene blocks with flanking SfiI sites (IDT DNA), and the SfiI fragment was subcloned into the modified pLV-EF1a-IRES-Puro-SfiI plasmid.
Countercurrent Chromatography:Article Title: A prenylated dsRNA sensor protects against severe COVID-19.
Article Snippet: .. Retroviral vectors and cell modification The lentiviral vector pSCRPSY (KT368137.1) has been previously described (5). pLV-EF1aIRES-Puro (Addgene plasmid #85132) or pLVEF1a-IRES-Blast (Addgene plasmid #85133) were modified by polymerase chain reaction (PCR) amplifying the TagRFP ORF (using pSCRPSY as template) flanked by directional SfiI sites, which were further flanked by BamHI and EcoRI restriction sites (forward oligo: 5′-CTC TCG GAT CCG GCC GAG AGG GCC ATG AGC GAG CTG ATT AAG-3′ and reverse oligo: 5′-CTC TCG AAT TCG GCC AGA GAG GCC TCA CTT GTG CCC CAG-3′), and the BamHI-EcoRI fragment was subcloned into the vectors to create the modified pLVEF1a-IRES-Puro-SfiI-TagRFP or pLV-EF1aIRES-Blast-SfiI-TagRFP constructs. .. The cDNA corresponding to the ORFs of the following OAS genes (GenBank accession number): OAS1p46 (NM_016816), human OAS3 (NM_006187), mouse Oas1A (NM_145211), bovine OAS1X (NM_178108), bovine OAS1Y (NM_001040606), bovine OAS1Z (AY650038), P. kuhlii (XM_036409709.1), C. dromedarius (XM_031443284), R. ferrumequinum OAS1 (short isoform: XM_033097132 / long isoform: ENSRFET00010016745), and P. alecto (NM_001290162) were synthesized as gene blocks with flanking SfiI sites (IDT DNA), and the SfiI fragment was subcloned into the modified pLV-EF1a-IRES-Puro-SfiI plasmid.
Construct:Article Title: A prenylated dsRNA sensor protects against severe COVID-19.
Article Snippet: .. Retroviral vectors and cell modification The lentiviral vector pSCRPSY (KT368137.1) has been previously described (5). pLV-EF1aIRES-Puro (Addgene plasmid #85132) or pLVEF1a-IRES-Blast (Addgene plasmid #85133) were modified by polymerase chain reaction (PCR) amplifying the TagRFP ORF (using pSCRPSY as template) flanked by directional SfiI sites, which were further flanked by BamHI and EcoRI restriction sites (forward oligo: 5′-CTC TCG GAT CCG GCC GAG AGG GCC ATG AGC GAG CTG ATT AAG-3′ and reverse oligo: 5′-CTC TCG AAT TCG GCC AGA GAG GCC TCA CTT GTG CCC CAG-3′), and the BamHI-EcoRI fragment was subcloned into the vectors to create the modified pLVEF1a-IRES-Puro-SfiI-TagRFP or pLV-EF1aIRES-Blast-SfiI-TagRFP constructs. .. The cDNA corresponding to the ORFs of the following OAS genes (GenBank accession number): OAS1p46 (NM_016816), human OAS3 (NM_006187), mouse Oas1A (NM_145211), bovine OAS1X (NM_178108), bovine OAS1Y (NM_001040606), bovine OAS1Z (AY650038), P. kuhlii (XM_036409709.1), C. dromedarius (XM_031443284), R. ferrumequinum OAS1 (short isoform: XM_033097132 / long isoform: ENSRFET00010016745), and P. alecto (NM_001290162) were synthesized as gene blocks with flanking SfiI sites (IDT DNA), and the SfiI fragment was subcloned into the modified pLV-EF1a-IRES-Puro-SfiI plasmid.
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