A Generally Applicable CRISPR/Cas9 Screening Technique to Identify Host Genes Required for Virus Infection as Applied to Human Cytomegalovirus (HCMV) Infection of Epithelial Cells

Methods Mol Biol. 2021:2244:247-264. doi: 10.1007/978-1-0716-1111-1_13.


Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 screens enable virus-host genetic screens to be undertaken in a more robust manner than previously possible and has had a tremendous impact in the field of virus study. Researchers can take advantage of the power of CRISPR genetic screens to discover virus-host interaction genes including host receptors and signaling molecules (Bazzone et al., mBio 10 (1): e02734-18, 2019; E et al., Proc Natl Acad Sci U S A 116(14):7043-7052, 2019; McDougall et al., Curr Opin Virol 29:87-100, 2018; Savidis et al., Cell Rep 16(1):232-246, 2016). In principle, lysis of cells late in the virus infection cycle allows one to screen for essential genes using pooled single-guide RNAs (sgRNAs) that collective target an entire host cell genome simply by identifying mutant cells that are resistant to virus-induced cell death. Here we focus on using this technique on epithelial cells to identify host targets required for human cytomegalovirus (HCMV) infection.

Keywords: CRISPR/Cas9 screening; HCMV; Human cytomegalovirus; Method; Primary epithelial cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • CRISPR-Cas Systems / genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Cytomegalovirus / genetics*
  • Epithelial Cells
  • Gene Editing / methods
  • Genetic Engineering / methods*
  • Genetic Testing / methods
  • Host Microbial Interactions / genetics*
  • Humans
  • Primary Cell Culture
  • RNA, Guide, Kinetoplastida / genetics
  • Virus Diseases / genetics


  • RNA, Guide