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Addgene inc sace2
Sace2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/no+149664/pm36094679-263-9-19?v=Addgene+inc
Average 93 stars, based on 3 article reviews

sace2 - by Bioz Stars, 2026-07

93/100 stars

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A Binding of monomeric sACE2‐8his proteins to Expi293F cells expressing S of BA.1 omicron was measured by flow cytometry. Data are mean ± SEM, N = 4 independent experiments. B Binding of dimeric sACE2 2 ‐IgG1 proteins was compared with the binding of antibodies authorized for therapeutic use in COVID‐19 patients. Binding to Expi293F cells expressing BA.1 omicron S was measured by flow cytometry. Data are mean ± SEM, N = 4 independent biological replicates. C Neutralization of BA.1 omicron pseudovirus. sACE2 2 ‐IgG1 (gray) or sACE2 2 .v2.4‐IgG1 (blue) were incubated with pseudovirus for 1 h before adding to HeLa‐hACE2‐11 cells. Infection 48 h later was measured by luciferase reporter gene expression. Data are mean ± SD, N = 3 independent biological replicates. D, E Authentic BA.1 omicron virus (isolate USA/MD‐HP20874/2021) was incubated with sACE2 2 ‐IgG1 (D) or sACE2 2 .v2.4‐IgG1 (E) for 1 h and added to Calu‐3 cells. Infection 48 h later was measured by RT–qPCR for the viral N gene. 3 μM remdesivir (black columns) is a positive neutralization control. Data are mean ± SD, N = 4 independent replicates.

Journal: EMBO Molecular Medicine

Article Title: An ACE2 decoy can be administered by inhalation and potently targets omicron variants of SARS‐CoV ‐2

doi: 10.15252/emmm.202216109

Figure Lengend Snippet: A Binding of monomeric sACE2‐8his proteins to Expi293F cells expressing S of BA.1 omicron was measured by flow cytometry. Data are mean ± SEM, N = 4 independent experiments. B Binding of dimeric sACE2 2 ‐IgG1 proteins was compared with the binding of antibodies authorized for therapeutic use in COVID‐19 patients. Binding to Expi293F cells expressing BA.1 omicron S was measured by flow cytometry. Data are mean ± SEM, N = 4 independent biological replicates. C Neutralization of BA.1 omicron pseudovirus. sACE2 2 ‐IgG1 (gray) or sACE2 2 .v2.4‐IgG1 (blue) were incubated with pseudovirus for 1 h before adding to HeLa‐hACE2‐11 cells. Infection 48 h later was measured by luciferase reporter gene expression. Data are mean ± SD, N = 3 independent biological replicates. D, E Authentic BA.1 omicron virus (isolate USA/MD‐HP20874/2021) was incubated with sACE2 2 ‐IgG1 (D) or sACE2 2 .v2.4‐IgG1 (E) for 1 h and added to Calu‐3 cells. Infection 48 h later was measured by RT–qPCR for the viral N gene. 3 μM remdesivir (black columns) is a positive neutralization control. Data are mean ± SD, N = 4 independent replicates.

Article Snippet: Plasmids for sACE2 2 ‐IgG1 (Addgene #154104), sACE2 2 .v2.4‐IgG1 (#154106), sACE2(18‐615)‐8his (#149268), sACE2(18‐615).v2.4‐8his (#149664), and Wuhan RBD‐8his (#145145) are available from Addgene.

Techniques: Binding Assay, Expressing, Flow Cytometry, Neutralization, Incubation, Infection, Luciferase, Gene Expression, Virus, Quantitative RT-PCR, Control

A Using flow cytometry, binding to Expi293F cells expressing S of BA.2 omicron was measured for monomeric sACE2‐8his proteins. Data are mean ± SEM, N = 3 independent biological replicates. B Binding of antibodies and dimeric sACE2 2 ‐IgG1 proteins to Expi293F cells expressing BA.2 omicron S measured by flow cytometry. Data are mean ± SEM, N = 4 independent experiments. C BA.2 omicron pseudovirus was incubated with sACE2 2 ‐IgG1 (gray) or sACE2 2 .v2.4‐IgG1 (blue) for 1 h and added to HeLa‐hACE2‐11 cells. Infection was measured after 48 h. Data are mean ± SD, N = 3 independent replicates. D, E Binding of monomeric sACE2‐8his proteins (D) and dimeric IgG1 proteins (E) to Expi293F cells expressing BA.4/BA.5 omicron S as measured by flow cytometry. Data are mean ± SEM, N = 3 independent experiments.

Journal: EMBO Molecular Medicine

Article Title: An ACE2 decoy can be administered by inhalation and potently targets omicron variants of SARS‐CoV ‐2

doi: 10.15252/emmm.202216109

Figure Lengend Snippet: A Using flow cytometry, binding to Expi293F cells expressing S of BA.2 omicron was measured for monomeric sACE2‐8his proteins. Data are mean ± SEM, N = 3 independent biological replicates. B Binding of antibodies and dimeric sACE2 2 ‐IgG1 proteins to Expi293F cells expressing BA.2 omicron S measured by flow cytometry. Data are mean ± SEM, N = 4 independent experiments. C BA.2 omicron pseudovirus was incubated with sACE2 2 ‐IgG1 (gray) or sACE2 2 .v2.4‐IgG1 (blue) for 1 h and added to HeLa‐hACE2‐11 cells. Infection was measured after 48 h. Data are mean ± SD, N = 3 independent replicates. D, E Binding of monomeric sACE2‐8his proteins (D) and dimeric IgG1 proteins (E) to Expi293F cells expressing BA.4/BA.5 omicron S as measured by flow cytometry. Data are mean ± SEM, N = 3 independent experiments.

Techniques: Flow Cytometry, Binding Assay, Expressing, Incubation, Infection

a – c , Based on simulations of RBD-bound ACE2, new polar interactions were identified between the ACE2.v2.4 (orange) and RBD (yellow) loops (red) ( a ). ACE2 mutations are presented as sticks colored cyan with nearby RBD residues as sticks colored yellow. New polar interactions between ACE2.v2.4 and RBD loops 1 ( b ) and 2 ( c ) are indicated by the dotted blue lines. d , MSM-weighted distance distributions of newly formed hydrogen bonds. For each interacting pair, the ACE2.v2.4 residue is listed first and the RBD residue is listed second. e , Probabilities for stable hydrogen bond interactions using a 4-Å distance criterion between accepter and donor. f , Root mean square fluctuation (RMSF) of RBD residues when bound to WT (cyan) or v2.4 (orange) ACE2 receptors. The RBD regions that interface with ACE2 are shaded gray. The absolute difference in RMSF between WT and v2.4 proteins is shown in black. Distance distributions, hydrogen bond probabilities and RMSF calculations were based on 40,000 frames from the simulations. Frames were selected based on the MSM stationary probability to represent the entire conformational ensemble. The error bars represent the 95% confidence intervals calculated from 20 bootstrapped samples.

Journal: Nature Chemical Biology

Article Title: Engineered ACE2 decoy mitigates lung injury and death induced by SARS-CoV-2 variants

doi: 10.1038/s41589-021-00965-6

Figure Lengend Snippet: a – c , Based on simulations of RBD-bound ACE2, new polar interactions were identified between the ACE2.v2.4 (orange) and RBD (yellow) loops (red) ( a ). ACE2 mutations are presented as sticks colored cyan with nearby RBD residues as sticks colored yellow. New polar interactions between ACE2.v2.4 and RBD loops 1 ( b ) and 2 ( c ) are indicated by the dotted blue lines. d , MSM-weighted distance distributions of newly formed hydrogen bonds. For each interacting pair, the ACE2.v2.4 residue is listed first and the RBD residue is listed second. e , Probabilities for stable hydrogen bond interactions using a 4-Å distance criterion between accepter and donor. f , Root mean square fluctuation (RMSF) of RBD residues when bound to WT (cyan) or v2.4 (orange) ACE2 receptors. The RBD regions that interface with ACE2 are shaded gray. The absolute difference in RMSF between WT and v2.4 proteins is shown in black. Distance distributions, hydrogen bond probabilities and RMSF calculations were based on 40,000 frames from the simulations. Frames were selected based on the MSM stationary probability to represent the entire conformational ensemble. The error bars represent the 95% confidence intervals calculated from 20 bootstrapped samples.

Article Snippet: Plasmids for the expression of myc-tagged human ACE2 (pCEP4-myc-ACE2, plasmid no. 141185; Addgene), 8 histidine-tagged monomeric sACE2 (ACE2 amino acids 1–615; pcDNA3-sACE2(WT)-8his, plasmid no. 149268 and pcDNA3-sACE2v2.4-8his, plasmid no. 149664; Addgene), SARS-CoV-1 S protein with C-terminal C9 tag (pcDNA3.1-SARS-spike, plasmid no. 145031; Addgene), 8his-tagged RBD (pcDNA3-SARS-CoV-2-S-RBD-8his, plasmid no. 145145; Addgene) and human IgG1-Fc fused dimeric sACE2 2 (ACE2 amino acids 1–732; pcDNA3-sACE2-WT(732)-IgG1, plasmid no. 154104 and pcDNA3-sACE2v2.4(732)-IgG1, plasmid no. 154106; Addgene) have been described previously.

Techniques: Residue

a , mRNA expression of ACE2 (NM_001371415.1) and TMPRSS2 (NM_001135099) in human lung epithelial A549 cells. A549 cells stably expressing hACE2 and hLMVECs were analyzed by one-step RT–PCR (top) and real-time PCR (bottom). Relative expression was normalized to glyceraldehyde 3-phosphate dehydrogenase expression. b , Cultured hACE2-A549 cells, A549 cells and hLMVECs were preincubated with sACE2 2 -IgG1 or sACE2 2 .v2.4-IgG1 at 5 µg ml −1 or 25 µg ml −1 for 1 h. SARS-CoV-2 pseudovirus (MOI = 0.1) was added to the cells, which were collected at 24 h. Virus entry was evaluated by luciferase activity. n = 4 replicates. c , A dose of 10 mg kg −1 sACE2 2 -IgG1, sACE2 2 .v2.4-IgG1 or buffer (PBS + 0.2% BSA) was administrated intravenously into K18-hACE2 transgenic mice for 30 min before SARS-CoV-2 pseudovirus (10 6 PFU) intraperitoneal injection. Tissue lysates were prepared at 24 h and virus entry in the selected organs was evaluated by luciferase activity. Buffer was applied as the control group, n = 4. d – g , K18-hACE2 transgenic mice were inoculated with the SARS-CoV-2 isolate WA-1/2020 at 1 × 10 4 PFU. Mice received control PBS or sACE2 2 .v2.4-IgG1 10 mg kg −1 via intravenous injection 12 h before inoculation. Mice were observed for survival ( d ) and body weight ( e ), n = 5. EBA as a marker of pulmonary endothelial permeability ( f ) and lung wet/dry ratio ( g ) as a measure of lung edema were quantified. Data are presented as the mean ± s.e.m., n = 4. b , c , f , g , P values were calculated by one-way ANOVA with Tukey post hoc test.

Journal: Nature Chemical Biology

Article Title: Engineered ACE2 decoy mitigates lung injury and death induced by SARS-CoV-2 variants

doi: 10.1038/s41589-021-00965-6

Figure Lengend Snippet: a , mRNA expression of ACE2 (NM_001371415.1) and TMPRSS2 (NM_001135099) in human lung epithelial A549 cells. A549 cells stably expressing hACE2 and hLMVECs were analyzed by one-step RT–PCR (top) and real-time PCR (bottom). Relative expression was normalized to glyceraldehyde 3-phosphate dehydrogenase expression. b , Cultured hACE2-A549 cells, A549 cells and hLMVECs were preincubated with sACE2 2 -IgG1 or sACE2 2 .v2.4-IgG1 at 5 µg ml −1 or 25 µg ml −1 for 1 h. SARS-CoV-2 pseudovirus (MOI = 0.1) was added to the cells, which were collected at 24 h. Virus entry was evaluated by luciferase activity. n = 4 replicates. c , A dose of 10 mg kg −1 sACE2 2 -IgG1, sACE2 2 .v2.4-IgG1 or buffer (PBS + 0.2% BSA) was administrated intravenously into K18-hACE2 transgenic mice for 30 min before SARS-CoV-2 pseudovirus (10 6 PFU) intraperitoneal injection. Tissue lysates were prepared at 24 h and virus entry in the selected organs was evaluated by luciferase activity. Buffer was applied as the control group, n = 4. d – g , K18-hACE2 transgenic mice were inoculated with the SARS-CoV-2 isolate WA-1/2020 at 1 × 10 4 PFU. Mice received control PBS or sACE2 2 .v2.4-IgG1 10 mg kg −1 via intravenous injection 12 h before inoculation. Mice were observed for survival ( d ) and body weight ( e ), n = 5. EBA as a marker of pulmonary endothelial permeability ( f ) and lung wet/dry ratio ( g ) as a measure of lung edema were quantified. Data are presented as the mean ± s.e.m., n = 4. b , c , f , g , P values were calculated by one-way ANOVA with Tukey post hoc test.

Techniques: Expressing, Stable Transfection, One Step RT-PCR, Real-time Polymerase Chain Reaction, Cell Culture, Virus, Luciferase, Activity Assay, Transgenic Assay, Injection, Control, Marker, Permeability

K18-hACE2 transgenic mice were inoculated with the SARS-CoV-2 isolate WA-1/2020 at 1 × 10 4 PFU. Group 1 received control PBS via intravenous injection 24 h postviral inoculation. Group 2 (v2.4 12 h) received sACE2 2 .v2.4-IgG1 10 mg kg −1 via intravenous injection 12 h postinoculation and then daily subsequent injections at the same dose. Group 3 (v2.4 24 h) received sACE2 2 .v2.4-IgG1 15 mg kg −1 via intravenous injection 24 h postinoculation and then daily subsequent injections at the same dose. a , b , Survival curves ( a ) and weights ( b ) for n = 10 mice for each group. c – e , Mouse lungs were obtained on day 7 postinoculation for assessment of lung vascular albumin permeability. c , Macroscopic images of lungs at baseline and day 7 postviral inoculation in three experimental groups without EBA (left) and with EBA injection (right). d , Quantification of EBA lung vascular endothelial permeability in all three experimental groups. e , Quantification of lung edema by wet/dry ratio in all three experimental groups at baseline and day 7 postinoculation. f , g , Time course of lung vascular endothelial permeability and edema formation of groups 2 (v2.4 12 h) ( f ) and 3 (v2.4 24 h) ( g ) as assessed by EBA assay and lung wet/dry ratio. Data are presented as the mean ± s.e.m. d , e , P values were calculated by two-way ANOVA with Tukey post hoc test. f , g , P values were calculated by one-way ANOVA with Tukey post hoc test.

Journal: Nature Chemical Biology

Article Title: Engineered ACE2 decoy mitigates lung injury and death induced by SARS-CoV-2 variants

doi: 10.1038/s41589-021-00965-6

Figure Lengend Snippet: K18-hACE2 transgenic mice were inoculated with the SARS-CoV-2 isolate WA-1/2020 at 1 × 10 4 PFU. Group 1 received control PBS via intravenous injection 24 h postviral inoculation. Group 2 (v2.4 12 h) received sACE2 2 .v2.4-IgG1 10 mg kg −1 via intravenous injection 12 h postinoculation and then daily subsequent injections at the same dose. Group 3 (v2.4 24 h) received sACE2 2 .v2.4-IgG1 15 mg kg −1 via intravenous injection 24 h postinoculation and then daily subsequent injections at the same dose. a , b , Survival curves ( a ) and weights ( b ) for n = 10 mice for each group. c – e , Mouse lungs were obtained on day 7 postinoculation for assessment of lung vascular albumin permeability. c , Macroscopic images of lungs at baseline and day 7 postviral inoculation in three experimental groups without EBA (left) and with EBA injection (right). d , Quantification of EBA lung vascular endothelial permeability in all three experimental groups. e , Quantification of lung edema by wet/dry ratio in all three experimental groups at baseline and day 7 postinoculation. f , g , Time course of lung vascular endothelial permeability and edema formation of groups 2 (v2.4 12 h) ( f ) and 3 (v2.4 24 h) ( g ) as assessed by EBA assay and lung wet/dry ratio. Data are presented as the mean ± s.e.m. d , e , P values were calculated by two-way ANOVA with Tukey post hoc test. f , g , P values were calculated by one-way ANOVA with Tukey post hoc test.

Techniques: Transgenic Assay, Control, Injection, Permeability

a – d , sACE2 carrying the v2.4 mutations has increased S binding compared with WT sACE2. Human Expi293F cells expressing myc-tagged S from 4 SARS-CoV-2 variants (Wuhan ( a ), B.1.1.7/Alpha ( b ), B1.351/Beta ( c ) and P.1/Gamma ( d )) were incubated with monomeric sACE2-8 h (black) or dimeric sACE2 2 -IgG1 (gray); bound protein was detected by flow cytometry. WT ACE2 proteins are shown as broken lines, v2.4 proteins are shown as solid lines. n = 3 independent replicates; data are shown as the mean ± s.e.m. e – i , Binding of sACE2 2 .v2.4-IgG1 is comparable to clinically effective mAbs. Binding of mAbs versus sACE2 2 .v2.4-IgG1 to the S proteins of SARS-CoV-2 VOCs (B.1.1.7/Alpha ( e ), B1.351/Beta ( f ), B.1.617.2/Delta ( g ) and P.1/Gamma ( h )) and S protein of SARS-CoV-1 ( i ), as measured by flow cytometry. n = 3 independent replicates; data are shown as the mean ± s.e.m.

Journal: Nature Chemical Biology

Article Title: Engineered ACE2 decoy mitigates lung injury and death induced by SARS-CoV-2 variants

doi: 10.1038/s41589-021-00965-6

Figure Lengend Snippet: a – d , sACE2 carrying the v2.4 mutations has increased S binding compared with WT sACE2. Human Expi293F cells expressing myc-tagged S from 4 SARS-CoV-2 variants (Wuhan ( a ), B.1.1.7/Alpha ( b ), B1.351/Beta ( c ) and P.1/Gamma ( d )) were incubated with monomeric sACE2-8 h (black) or dimeric sACE2 2 -IgG1 (gray); bound protein was detected by flow cytometry. WT ACE2 proteins are shown as broken lines, v2.4 proteins are shown as solid lines. n = 3 independent replicates; data are shown as the mean ± s.e.m. e – i , Binding of sACE2 2 .v2.4-IgG1 is comparable to clinically effective mAbs. Binding of mAbs versus sACE2 2 .v2.4-IgG1 to the S proteins of SARS-CoV-2 VOCs (B.1.1.7/Alpha ( e ), B1.351/Beta ( f ), B.1.617.2/Delta ( g ) and P.1/Gamma ( h )) and S protein of SARS-CoV-1 ( i ), as measured by flow cytometry. n = 3 independent replicates; data are shown as the mean ± s.e.m.

Techniques: Binding Assay, Expressing, Incubation, Flow Cytometry

The 3 groups of K18-hACE2 transgenic mice were inoculated with the SARS-CoV-2 variant P.1 (Brazil) at 1 × 10 4 PFU. Group 1 (PBS): PBS was given by intravenous injection 24 h postinoculation. Group 2 (v2.4 12 h): sACE2 2 .v2.4-IgG1 10 mg kg −1 was given by intravenous injection 12 h postinoculation. Group 3 (v2.4 24 h): sACE2 2 .v2.4-IgG1 15 mg kg −1 was given by intravenous injection 24 h postinoculation. a , b , Mice were injected once per day for 7 d. Survival probability was calculated ( a ) and mouse weights were measured ( b ). n = 10 mice for each group. c , d , Mouse lungs were examined on day 6 postinoculation to evaluate lung vascular permeability and lung edema using the EBA assay ( c ) and lung wet/dry ratio ( d ), with baseline mouse lungs as controls. e , Viral loads of SARS-CoV-2 in the lungs obtained at baseline and day 6 postinoculation of the SARS-CoV-2 variant P.1 (Brazil) were measured by real-time qPCR for mRNA expression of SARS-CoV-2 S protein and SARS-CoV-2 NSP. f , A viral plaque formation assay was performed to measure the viral loads of SARS-CoV-2 in lungs obtained at baseline and on day 6 postinoculation of the SARS-CoV-2 P.1 (Brazil) variant. c – e , f , n = 4. Data are presented as the mean ± s.e.m. c , d , P values were calculated by two-way ANOVA with Tukey post hoc test. e , f , P values were calculated by one-way ANOVA with Tukey post hoc test.

Journal: Nature Chemical Biology

Article Title: Engineered ACE2 decoy mitigates lung injury and death induced by SARS-CoV-2 variants

doi: 10.1038/s41589-021-00965-6

Figure Lengend Snippet: The 3 groups of K18-hACE2 transgenic mice were inoculated with the SARS-CoV-2 variant P.1 (Brazil) at 1 × 10 4 PFU. Group 1 (PBS): PBS was given by intravenous injection 24 h postinoculation. Group 2 (v2.4 12 h): sACE2 2 .v2.4-IgG1 10 mg kg −1 was given by intravenous injection 12 h postinoculation. Group 3 (v2.4 24 h): sACE2 2 .v2.4-IgG1 15 mg kg −1 was given by intravenous injection 24 h postinoculation. a , b , Mice were injected once per day for 7 d. Survival probability was calculated ( a ) and mouse weights were measured ( b ). n = 10 mice for each group. c , d , Mouse lungs were examined on day 6 postinoculation to evaluate lung vascular permeability and lung edema using the EBA assay ( c ) and lung wet/dry ratio ( d ), with baseline mouse lungs as controls. e , Viral loads of SARS-CoV-2 in the lungs obtained at baseline and day 6 postinoculation of the SARS-CoV-2 variant P.1 (Brazil) were measured by real-time qPCR for mRNA expression of SARS-CoV-2 S protein and SARS-CoV-2 NSP. f , A viral plaque formation assay was performed to measure the viral loads of SARS-CoV-2 in lungs obtained at baseline and on day 6 postinoculation of the SARS-CoV-2 P.1 (Brazil) variant. c – e , f , n = 4. Data are presented as the mean ± s.e.m. c , d , P values were calculated by two-way ANOVA with Tukey post hoc test. e , f , P values were calculated by one-way ANOVA with Tukey post hoc test.

Techniques: Transgenic Assay, Variant Assay, Injection, Permeability, Expressing, Plaque Formation Assay

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