plasmid pk18 hace21  (Addgene inc)

Journal: Microbes and Infection

Article Title: Mice Humanized for MHC and hACE2 with High Permissiveness to SARS-CoV-2 Omicron Replication

doi: 10.1016/j.micinf.2023.105142

Figure Lengend Snippet: Permissiveness of HHD-DR1.ACE2 Hu1 and HHD-DR1.ACE2 Hu2 transgenic mice to SARS-CoV-2 replication. hACE2 + offspring from the founders #8, #9 (HHD-DR1.ACE2 Hu1 ) or #11 (HHD-DR1.ACE2 Hu2 ), resulting from a conventional DNA-based transgenesis, or B6.K18-hACE2 IP-THV positive control mice were inoculated i.n. with 0.3 × 10 5 TCID 50 /mouse of SARS-CoV-2 Delta variant. A. Lung and brain viral RNA contents were determined by E- (top) or sub-genomic Esg- (bottom) specific qRT-PCR at 4 dpi. Red lines indicate the qRT-PCR detection limits. B. Quantification of hACE-2 mRNA in the lung and brain of the same mice. Statistical significance was evaluated by one-way Anova (*= p < 0.05, **= p < 0.01, ***= p < 0.001).

Article Snippet: The pK18- hACE2 plasmid, purchased from Addgene (Watertown, MA), was double digested with Hpa1 and Xba1 enzymes, and the 6.8 kb DNA fragment generated was purified with Qiaex II gel extraction kit (Qiagen).

Techniques: Transgenic Assay, Positive Control, Variant Assay, Quantitative RT-PCR

Journal: Microbes and Infection

Article Title: Mice Humanized for MHC and hACE2 with High Permissiveness to SARS-CoV-2 Omicron Replication

doi: 10.1016/j.micinf.2023.105142

Figure Lengend Snippet: Permissiveness of HHD-DR1.ACE2 Hu3 transgenic mice to SARS-CoV-2 replication and inflammatory status of their lungs and brain after SARS-CoV-2 inoculation. hACE2 + offspring from the founders #30 or #47 (HHD-DR1.ACE2 Hu3 ), resulting from a lentiviral vector-based transgenesis, were inoculated i.n. with 0.3 × 10 5 TCID 50 /mouse of SARS-CoV-2 Delta variant. A. Lung and brain viral RNA contents were determined by E-specific or sub-genomic Esg-specific qRT-PCR at 4 dpi. B. Quantification of hACE-2 mRNA in the lung and brain of the same mice. C. Heatmaps represent log 2 fold change in cytokine and chemokine mRNA expression in the lungs or brain of the same mice ( n = 4-5/group). Data were normalized versus untreated controls. Statistical significance was evaluated by Mann-Whitney test (ns = not significant, * = p < 0.05, ** = p < 0.01). D. Pearson correlation coefficient of the analytes studied in the brain of SARS-CoV-2 Delta-inoculated HHD-DR1.ACE2 Hu3 mice.

Article Snippet: The pK18- hACE2 plasmid, purchased from Addgene (Watertown, MA), was double digested with Hpa1 and Xba1 enzymes, and the 6.8 kb DNA fragment generated was purified with Qiaex II gel extraction kit (Qiagen).

Techniques: Transgenic Assay, Plasmid Preparation, Variant Assay, Quantitative RT-PCR, Expressing, MANN-WHITNEY

Journal: Microbes and Infection

Article Title: Mice Humanized for MHC and hACE2 with High Permissiveness to SARS-CoV-2 Omicron Replication

doi: 10.1016/j.micinf.2023.105142

Figure Lengend Snippet: Permissiveness of HHD-DR1.ACE2 Hu1/2/3 mice to replication of SARS-CoV-2 Omicron BA.1 or BA.5 sub-variants. HHD-DR1.ACE2 Hu1/2/3 mice or B6.K18-hACE2 IP-THV positive control mice were inoculated i.n. with 0.3 × 10 5 TCID 50 /mouse of SARS-CoV-2 Omicron BA.1 or BA.5. A. Lung and B. brain viral RNA contents were determined by E-specific (top) or sub-genomic Esg-specific (bottom) qRT-PCR at 4 dpi with Omicron BA.1. C-D. Comparative quantification of hACE-2 mRNA in the lung and brain of individual B6.K18-hACE2 IP-THV or HHD-DR1.ACE2 Hu1/2/3 transgenic mice. E-F. Lung and brain viral RNA contents were determined by E-specific (top) or sub-genomic Esg-specific (bottom) qRT-PCR at 4 dpi with Omicron BA.5. Statistical significance was evaluated by one-way Anova (*= p < 0.05, **= p < 0.01). ND = Not determined.

Article Snippet: The pK18- hACE2 plasmid, purchased from Addgene (Watertown, MA), was double digested with Hpa1 and Xba1 enzymes, and the 6.8 kb DNA fragment generated was purified with Qiaex II gel extraction kit (Qiagen).

Techniques: Positive Control, Quantitative RT-PCR, Transgenic Assay

Journal: bioRxiv

Article Title: Mice Humanized for Major Histocompatibility Complex and Angiotensin-Converting Enzyme 2 with High Permissiveness to SARS-CoV-2 Omicron Replication

doi: 10.1101/2022.12.01.518541

Figure Lengend Snippet: hACE2 + offspring from the founders #8, #9 (HHD-DR1.ACE2 Hu1 ) or #11 (HHD-DR1.ACE2 Hu2 ), resulting from a conventional DNA-based transgenesis, or B6.K18-hACE2 IP-THV positive control mice were inoculated i.n. with 0.3 × 10 5 TCID 50 /mouse of SARS-CoV-2 Delta variant. A Lung and brain viral RNA contents were determined by E-(top) or sub-genomic Esg-(bottom) specific qRT-PCR at 4 dpi. Red lines indicate the qRT-PCR detection limits. B Quantification of hACE-2 mRNA in the lung and brain of the same mice. Statistical significance was evaluated by Mann-Whitney test (*= p < 0.05, **= p < 0.01, ***= p < 0.001).

Article Snippet: Zygotes from HHD-DR1 mice were microinjected with a 6.8-kb Hpal I-XbaI fragment from the pK18-hACE2 vector at 2 ng/μl ( McCray et al ., 2007 ), purchased from Addgene (Watertown, MA).

Techniques: Positive Control, Variant Assay, Quantitative RT-PCR, MANN-WHITNEY

Journal: bioRxiv

Article Title: Mice Humanized for Major Histocompatibility Complex and Angiotensin-Converting Enzyme 2 with High Permissiveness to SARS-CoV-2 Omicron Replication

doi: 10.1101/2022.12.01.518541

Figure Lengend Snippet: hACE2 + offspring from the founders #30 or #47 (HHD-DR1.ACE2 Hu3 ), resulting from a lentiviral vector-based transgenesis, were inoculated i.n. with 0.3 × 10 5 TCID 50 /mouse of SARS-CoV-2 Delta variant. A-B Lung (A) and brain (B) viral RNA contents were determined by E-specific (top) or sub-genomic Esg-specific (bottom) qRT-PCR at 4 dpi. C-D Quantification of hACE-2 mRNA in the lung and brain of the same mice. Statistical significance was evaluated by Mann-Whitney test (*= p < 0.05, **= p < 0.01).

Article Snippet: Zygotes from HHD-DR1 mice were microinjected with a 6.8-kb Hpal I-XbaI fragment from the pK18-hACE2 vector at 2 ng/μl ( McCray et al ., 2007 ), purchased from Addgene (Watertown, MA).

Techniques: Plasmid Preparation, Variant Assay, Quantitative RT-PCR, MANN-WHITNEY