Knockout of cGAS and STING Rescues Virus Infection of Plasmid DNA-Transfected Cells

J Virol. 2015 Nov;89(21):11169-73. doi: 10.1128/JVI.01781-15. Epub 2015 Aug 26.

Abstract

It is well known that plasmid DNA transfection, prior to virus infection, negatively affects infection efficiency. Here, we show that cytosolic plasmid DNA activates the cGAS/STING signaling pathway, which ultimately leads to the induction of an antiviral state of the cells. Using a transient one-plasmid clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system, we generated cGAS/STING-knockout cells and show that these cells can be infected after plasmid DNA transfection as efficiently as nontransfected cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems
  • Coxsackievirus Infections / metabolism*
  • Cytosol / metabolism*
  • Flow Cytometry
  • Gene Knockout Techniques
  • Gene Transfer Techniques*
  • Green Fluorescent Proteins
  • HeLa Cells
  • Humans
  • Luminescent Proteins
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mengovirus / metabolism
  • Nucleotidyltransferases / genetics
  • Nucleotidyltransferases / metabolism*
  • Plasmids / genetics
  • Plasmids / metabolism*
  • Red Fluorescent Protein
  • Signal Transduction / genetics
  • Signal Transduction / physiology*
  • Transfection / methods

Substances

  • Luminescent Proteins
  • Membrane Proteins
  • STING1 protein, human
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Nucleotidyltransferases
  • cGAS protein, human