Journal
VIRUSES-BASEL
Volume 13, Issue 5, Pages -Publisher
MDPI
DOI: 10.3390/v13050783
Keywords
KSHV; CRISPR-interference; dCas9-KRAB; Kaposi's sarcoma-associated herpesvirus; gene expression; gene silencing
Categories
Funding
- UC Cancer Research Coordinating Committee [CTN-19-586108]
- National Science Foundation California LSAMP Bridge [HRD-1701365]
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CRISPR interference (CRISPRi) can efficiently silence latent and lytic viral genes of Kaposi's sarcoma-associated herpesvirus (KSHV) in epithelial and B-cells, providing a functional genomics tool for studying virus-host interactions.
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 similar to 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.
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