Generation of a VeroE6 Pgp gene knock out cell line and its use in SARS-CoV-2 antiviral study

• DOI: 10.1016/j.antiviral.2022.105429
• Publication Date: 2022-10-05T15:23:06Z
• Authors: Yuao Zhu; Joe Binder; Irina Yurgelonis; Devendra K. Rai; Sarah Lazarro; Chester Costales; Keith Kobylarz; Patricia McMonagle; Claire M. Steppan; Lisa Aschenbrenner; Annaliesa S. Ande...; Rhonda D. Cardin
• Keywords: Vero cell; Calcein; Antiviral drug; Efflux

Vero cells are widely used for antiviral tests and virology research SARS-CoV-2 as well viruses from various other families. However, generally express high levels of multi-drug resistance 1 (MDR1) or Pgp protein, the efflux transporter foreign substances including many compounds, affecting activity interpretation data. To address this, a gene knockout VeroE6 cell line (VeroE6-Pgp-KO) was generated using CRISPR-CAS9 technology. These no longer expressed protein indicated by flow cytometry analysis following staining with Pgp-specific monoclonal antibody. They also showed significantly reduced in calcein acetoxymethyl ester (calcein AM) assay. The VeroE6-Pgp-KO parental were each infected to test activities remdesivir nirmatrelvir, two known substrates, presence absence inhibitor. compounds similar that observed Thus, newly established adds new vitro virus infection system possibly therapies without need control activity. Removal inhibitor assays will lead less data variation prevent failed assays.