Overexpression of human ACE2 protein in mouse fibroblasts stably transfected with the intact ACE2 gene

Teng, Feiyue; Gang, Oleg; Freimuth, Paul

doi:10.1016/j.virol.2024.109988

Overexpression of human ACE2 protein in mouse fibroblasts stably transfected with the intact ACE2 gene

Journal Article · Wed Jan 31 00:00:00 EST 2024 · Virology

DOI:https://doi.org/10.1016/j.virol.2024.109988· OSTI ID:2338131

Teng, Feiyue ^[1]; Gang, Oleg ^[2]; ^[1]

Brookhaven National Laboratory (BNL), Upton, NY (United States)
Brookhaven National Laboratory (BNL), Upton, NY (United States); Columbia Univ., New York, NY (United States)

Infection by SARS-CoV-2 is dependent on binding of the viral spike protein to angiotensin converting enzyme 2 (ACE2), a membrane glycoprotein expressed on epithelial cells in the human upper respiratory tract. Recombinant ACE2 protein has potential application for anti-viral therapy. Here we co-transfected mouse fibroblasts (A9 cells) with a cloned fragment of human genomic DNA containing the intact ACE2 gene and an unlinked neomycin phosphotransferase gene, and then selected stable neomycin-resistant transfectants. Transfectant clones expressed ACE2 protein at levels that were generally proportional to the number of ACE2 gene copies integrated in the cell genome, ranging up to approximately 50 times the level of ACE2 present of Vero-E6 cells. Cells overexpressing ACE2 were hypersensitive to infection by spike-pseudotyped vesicular stomatitis virus (VSV-S), and adsorption of VSV-S to these cells occurred at an accelerated rate compared to Vero-E6 cells. In conclusion, the transfectant cell clones described here therefore have favorable attributes as feedstocks for large-scale production of recombinant human ACE2 protein.

View Accepted Manuscript (DOE)

Research Organization:: Brookhaven National Laboratory (BNL), Upton, NY (United States)

Sponsoring Organization:: USDOE Laboratory Directed Research and Development (LDRD) Program; USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities (SUF)

Grant/Contract Number:: SC0012704

OSTI ID:: 2338131

Report Number(s):: BNL--225504-2024-JAAM

Journal Information:: Virology, Journal Name: Virology Vol. 592; ISSN 0042-6822

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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77 NANOSCIENCE AND NANOTECHNOLOGY
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Overexpression of human ACE2 protein in mouse fibroblasts stably transfected with the intact ACE2 gene

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References (49)

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