Application of pseudotyped virus particles to monitor Ebola virus and SARS-CoV-2 viral entry in human cell lines

STAR Protoc. 2021 Dec 17;2(4):100818. doi: 10.1016/j.xpro.2021.100818. Epub 2021 Aug 27.

Abstract

Experimental work on highly pathogenic viruses such as Ebola virus (EBOV) and severe acute respiratory syndrome coronavirus-2 requires high-level biosafety facilities. Here, we provide a detailed step-by-step protocol which details the production and application of replication-incompetent murine leukemia virus-based pseudotyped particles to monitor and quantify the viral entry efficiency in human cell lines under biosafety level-2 conditions. We describe the use of viral particles encoding luciferase gene and the quantification of transduction efficiency by measuring luciferase activity. For complete details on the use and execution of this protocol, please refer to Imre et al. (2021).

Keywords: Cell culture; Cell-based Assays; Immunology; Microbiology; Molecular Biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 / diagnosis*
  • COVID-19 / virology
  • Ebolavirus / physiology*
  • HEK293 Cells
  • Hemorrhagic Fever, Ebola / diagnosis*
  • Hemorrhagic Fever, Ebola / virology
  • Humans
  • SARS-CoV-2 / physiology*
  • Viral Pseudotyping / methods*
  • Virion
  • Virus Internalization*