MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT ACE2 TMPRSS2 RECEPTOR
1 vial (2 x 106) frozen cells
Cellbanker 2 (Amsbio 11891)
Expression vector containing full-length human ACE2 cDNA (GenBank Accession Number: NM_021804.1) w ith FLAG tag sequence at N-terminus and expression vector containing full-length human TMPRSS2 cDNA (GenBank Accession Number: NM_005656.3) with HA tag sequence at C-terminus expression vector
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 5 ug/ml blasticidin, 250 ug/mL hygromycin
Background: Angiotensin converting enzyme (ACE2) receptors locate in the human oral pharynx and upper airway where the high replication rate of Corona virus happens more easily. ACE2 has been confirmed as the cause of SARS-CoV-2 internalization in concert with TMPRSS2 (transmembrane protease serin 2). TMPRSS2 facilitates ACEs cell entry by priming the spike S protein of the virus. Therapeutic strategies that focus on the biology of ACE2 and TMPRSS2 may become beneficial for the treatment of lung disease.
Application: Functional assays
Figure 1. Dose-dependent inhibition of protease activity, measured w ith fluorogenic peptide substrate specific to ACE2 (a) and TMPRSS2 (b) Figure 2. Receptor expression on cell surface measured by flow cytometry (FACS) using anti-Flag antibody (a) and primary antibody Rb mAb to HA-tag and secondary antibody R- Phycoerythrin conjugate (b). Thin line: parental cells; thick line: receptor-expressing cells.
References: Liu et al. (2020) The science underlying COVID-19, Implications for the Cardiovascular System. Circulation. 2020;142:68–78. DOI: 10.1161/CIRCULATIONAHA.120.047549
Hoffmann et al. (2020) SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 is Blocked by a Clinically Proven Protease Inhibitor. Cell 181, 271–280, April 16, 2020
Zhang, et al. Angiotensin- converting enzyme 2 (ACE2) as a SARS- CoV-2 receptor: molecular mechanisms and potential therapeutic target. (2020). Intensive Care Med., 46, 586–5905.