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性能数据
图1. GeneCopoeia稳定细胞系中ACE2和TMPRSS2的相对mRNA表达水平。利用对每个基因特异的引物，通过RT-qPCR扩增mRNA。ACE2和TMPRSS2的Ct值被归一化为管家基因GAPDH的Ct值。相对表达水平根据2^ΔΔCt计算。A. SL221，ACE2 mRNA在亲本细胞系HEK293T和过表达HEK293T细胞系中的表达。B. SL222，ACE2和TMPRSS2 mRNA在亲本细胞系HEK293T中的表达和ACE2-和TMPRSS2-过表达细胞系。
图2. 表达ACE2的HEK293T细胞（Cat.#SL221）、表达ACE2/TMPRSS2的HEH293T细胞和HEK293T细胞的细胞表面上ACE2蛋白表达的免疫荧光（IF）分析。将未渗透的细胞固定、阻断并与抗ACE2的一抗孵育，然后洗涤，并与有荧光标记的二抗孵育，再次洗涤并用DAPI染色。
图3. 表达TMPRSS2的HEK293T细胞（Cat.#SL222）和HEK293T细胞的细胞表面上TMPRSS2蛋白表达的免疫荧光（IF）分析。将未渗透的细胞固定、阻断并与抗TMPRSS2的一抗孵育，然后洗涤，并与有荧光标记的二抗孵育，再次洗涤并用DAPI染色。
GeneCopoeia的研发团队测试了SARS-CoV-2 S 蛋白假慢病毒感染ACE2表达细胞的能力，如下图4所示。A.
B.
C.
图4. 用表达eGFP/hLuc的SARS-CoV-2 S假慢病毒（假病毒）转导表达ACE2-或ACE2/TMPRSS2的HEK293T细胞。 用10 μl假病毒在96孔板中转导每个细胞系。转导72小时后，在荧光显微镜下对细胞进行拍照，随后测量荧光素酶活性或进行FACS分选。SL221：表达ACE2的HEK293T细胞。SL222：表达ACE2和TMPRSS2的HEK293T细胞。SP101：全长S蛋白，D614变体。SP103：全长S蛋白，D614G变体。SP001：截短S蛋白，D614变体。SP003：截短S蛋白，D614G变体。A. 荧光显微镜。B. 荧光素酶活性测定。使用GeneCopoeia的Luc-Pair™ Firefly Luciferase HS Assay Kit（货号LF007），从假病毒转导细胞培养物中提取20 μl细胞裂解液，分析萤火虫荧光素酶活性，用FACS分选法测定转导的单位滴度units/ml。
资源 在线研讨会
标题:来自GeneCopoeia的COVID-19冠状病毒研究的先进工具
2020年9月9日，星期三
由SARS-CoV-2冠状病毒引起的新冠肺炎全球大流行凸显了对诊断试剂、抗病毒治疗和疫苗开发的迫切需求。在本次网络研讨会中，我们将讨论GeneCopoeia先进的解决方案，包括检测试剂盒、SARS-CoV-2假病毒以及研究对抗这种致命疾病所需的其他工具。
观看录制的网络研讨会/下载 幻灯片
已发表的文章
查看近期使用GeneCopoeia SARS-CoV-2 S蛋白假慢病毒产品的已发表文章
2023
Kuroda, T., et al. (2023). Efficacy comparison of 3CL protease inhibitors ensitrelvir and nirmatrelvir against SARS-CoV-2 in vitro and in vivo. J Antimicrob Chemother doi: 10.1093/jac/dkad027 [ACE2 and TMPRSS2-expressing HEK293T cells ]
Osaka, T., et al. (2023). Cross-Reactivity of Antibodies in Intravenous Immunoglobulin Preparation for Protection against SARS-CoV-2. Microorganisms doi: 10.3390/microorganisms11020471 [ACE2- and TMPRSS2-expressing HEK293T stable cell line ]
Chan, J.F-K., et al. (2023). Altered host protease determinants for SARS-CoV-2 Omicron. Sci Adv doi: 10.1126/sciadv.add3867 [ACE2-expressing HEK293T stable cell line ]
Merluza, J., et al. (2023). Validation and Establishment of the SARS-CoV-2 Lentivirus Surrogate Neutralization Assay as a Prescreening Tool for the Plaque Reduction Neutralization Test. Microbiol Spectr doi: 10.1128/spectrum.03789-22 [ACE2- and TMPRSS2-expressing HEK293T stable cell line ]
Nakata, Y., et al. (2023). Cellular APOBEC3A deaminase drives mutations in the SARS-CoV-2 genome. Nucleic Acids Res doi: 10.1093/nar/gkac1238 [ACE2- and TMPRSS2-expressing HEK293T stable cell line ]
2022
Sun, P.F., et al. (2022). Asymptomatic or symptomatic SARS-CoV-2 infection plus vaccination confers increased adaptive immunity to variants of concern. iScience doi: 10.1016/j.isci.2022.105202 [HEK293-ACE2 expressing cell lines ]
Suzuki, Y., et al. (2022). Design and lyophilization of lipid nanoparticles for mRNA vaccine and its robust immune response in mice and nonhuman primates. Mol Ther Nucleic Acids doi: 10.1016/j.omtn.2022.09.017 [ACE2 and TMPRSS2-expressing HEK293T cells ]
Hwang, J-Y., et al. (2022). Humoral and Cellular Responses to COVID-19 Vaccines in SARS-CoV-2 Infection-Naïve and -Recovered Korean Individuals . Vaccines doi: 10.3390/vaccines10020332 [ACE2-expressing HEK293T stable cell line ]
Park, H., et al. (2022). TMED3 Complex Mediates ER Stress-Associated Secretionof CFTR, Pendrin, and SARS-CoV-2 Spike . Advanced Science doi: 10.1002/advs.202105320 [ACE2- and TMPRSS2-expressing HEK293T stable cell line ]
Sim, J-R., et al. (2022). Amelioration of SARS-CoV-2 infection by ANO6 phospholipid scramblase inhibition . Cell Reports doi: 10.1016/j.celrep.2022.111117 [ACE2- and TMPRSS2-expressing HEK293T stable cell line ]
Grodzki, M., et al. (2022). Genome-scale CRISPR screens identify host factors that promote human coronavirus infection . Genome Medicine doi: 10.1186/s13073-022-01013-1 [ACE2-expressing HEK293T stable cell line ]
Ariumi, Y. (2022). Host Cellular RNA Helicases Regulate SARS-CoV-2 Infection . Journal of Virology doi: 10.1128/jvi.00002 [ACE2-expressing HEK293T stable cell line ]
Pivniouk, V., et al. (2022). The OM-85 bacterial lysate inhibits SARS-CoV-2 infection of epithelial cells by downregulating SARS-CoV-2 receptor expression . Journal of Allergy and Clinical Immunology doi: 10.1016/j.jaci.2021.11.019 [ACE2-expressing HEK293T stable cell line ]
Travis, B.J., et al. (2022). Significance of chlorine-dioxide-based oral rinses in preventing SARS-CoV-2 cell entry. Oral Diseases doi: 10.1111/odi.14319 [ACE2-expressing HEK293T stable cell line ]
2021
Hashizume, M., et al. (2021). Population-Specific ACE2 Single-Nucleotide Polymorphisms Have Limited Impact on SARS-CoV-2 Infectivity In Vitro . Viruses doi: 10.3390/v13010067 [ACE2-expressing HEK293T stable cell line ]
Ao, Z., et al .(2021). Identification and evaluation of the inhibitory effect of Prunella vulgaris extract on SARS-coronavirus 2 virus entry . PLOS ONE doi: 10.1371/journal.pone.0251649 [ACE2-expressing HEK293T stable cell line ]
Paidi, R.K., et al. (2021). Eugenol, a Component of Holy Basil (Tulsi) and Common Spice Clove, Inhibits the Interaction Between SARS-CoV-2 Spike S1 and ACE2 to Induce Therapeutic Responses . Journal of Neuroimmune Pharmacology doi: 10.1007/s11481-021-10028-1 [ACE2-expressing HEK293T stable cell line ]
Paidi, R.K., et al. (2021). Selective Inhibition of the Interaction between SARS-CoV-2 Spike S1 and ACE2 by SPIDAR Peptide Induces Anti-Inflammatory Therapeutic Responses . Journal of Immunology doi: 10.4049/jimmunol.2100144 [ACE2-expressing HEK293T stable cell line ]
2020
