CRISPR Interference Efficiently Silences Latent and Lytic Viral Genes in Kaposi's Sarcoma-Associated Herpesvirus-Infected Cells

Viruses. 2021 Apr 28;13(5):783. doi: 10.3390/v13050783.

Abstract

Uncovering viral gene functions requires the modulation of gene expression through overexpression or loss-of-function. CRISPR interference (CRISPRi), a modification of the CRISPR-Cas9 gene editing technology, allows specific and efficient transcriptional silencing without genetic ablation. CRISPRi has been used to silence eukaryotic and prokaryotic genes at the single-gene and genome-wide levels. Here, we report the use of CRISPRi to silence latent and lytic viral genes, with an efficiency of ~80-90%, in epithelial and B-cells carrying multiple copies of the Kaposi's sarcoma-associated herpesvirus (KSHV) genome. Our results validate CRISPRi for the analysis of KSHV viral elements, providing a functional genomics tool for studying virus-host interactions.

Keywords: CRISPR-interference; KSHV; Kaposi’s sarcoma-associated herpesvirus; dCas9-KRAB; gene expression; gene silencing.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Line
  • Cells, Cultured
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Gene Editing*
  • Gene Expression Regulation, Viral
  • Gene Silencing*
  • Genes, Reporter
  • Genes, Viral
  • Herpesviridae Infections / virology
  • Herpesvirus 8, Human / genetics*
  • Humans
  • RNA, Guide, CRISPR-Cas Systems
  • Virus Activation / genetics*
  • Virus Latency / genetics*

Substances

  • RNA, Guide, CRISPR-Cas Systems