Suppression of Epstein-Barr virus DNA load in latently infected nasopharyngeal carcinoma cells by CRISPR/Cas9

Kit-San Yuen; Zhong-Min Wang; Nok-Hei Mickey Wong; Zhi-Qian Zhang; Tsz-Fung Cheng; Wai-Yin Lui; Chi-Ping Chan; Dong-Yan Jin

doi:10.1016/j.virusres.2017.04.019

Suppression of Epstein-Barr virus DNA load in latently infected nasopharyngeal carcinoma cells by CRISPR/Cas9

Virus Res. 2018 Jan 15:244:296-303. doi: 10.1016/j.virusres.2017.04.019. Epub 2017 Apr 27.

Authors

Kit-San Yuen¹, Zhong-Min Wang¹, Nok-Hei Mickey Wong¹, Zhi-Qian Zhang¹, Tsz-Fung Cheng¹, Wai-Yin Lui¹, Chi-Ping Chan¹, Dong-Yan Jin²

Affiliations

¹ School of Biomedical Sciences, The University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong.
² School of Biomedical Sciences, The University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong. Electronic address: dyjin@hku.hk.

PMID: 28456574
DOI: 10.1016/j.virusres.2017.04.019

Abstract

Epstein-Barr virus (EBV) infects more than 90% of the world's adult population. Once established, latent infection of nasopharyngeal epithelial cells with EBV is difficult to eradicate and might lead to the development of nasopharyngeal carcinoma (NPC) in a small subset of individuals. In this study we explored the anti-EBV potential of CRISPR/Cas9 targeting of EBV genome in infected NPC cells. We designed gRNAs to target different regions of the EBV genome and transfected them into C666-1 cells. The levels of EBV DNA in transfected cells were decreased by about 50%. The suppressive effect on EBV DNA load lasted for weeks but could not be further enhanced by re-transfection of gRNA. Suppression of EBV by CRISPR/Cas9 did not affect survival of C666-1 cells but sensitized them to chemotherapeutic killing by cisplatin and 5-fluorouracil. Our work provides the proof-of-principle for suppressing EBV DNA load with CRISPR/Cas9 and a potential new strategy to sensitize EBV-infected NPC cells to chemotherapy.

Keywords: CRISPR/Cas9; EBNA1; Epstein-Barr virus; Nasopharyngeal carcinoma.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antineoplastic Agents / pharmacology
Bacterial Proteins / genetics*
Bacterial Proteins / metabolism
CRISPR-Associated Protein 9
CRISPR-Cas Systems*
Cell Line, Tumor
Cell Survival / drug effects
Cisplatin / pharmacology
Clustered Regularly Interspaced Short Palindromic Repeats
DNA, Viral / genetics*
DNA, Viral / metabolism
Endonucleases / genetics*
Endonucleases / metabolism
Epithelial Cells / drug effects
Epithelial Cells / pathology
Epithelial Cells / virology
Fluorouracil / pharmacology
Gene Editing / methods*
Genome, Viral*
Herpesvirus 4, Human / drug effects
Herpesvirus 4, Human / genetics*
Herpesvirus 4, Human / growth & development
Herpesvirus 4, Human / metabolism
Humans
Nasopharynx / drug effects
Nasopharynx / pathology
Nasopharynx / virology
Plasmids / chemistry
Plasmids / metabolism
RNA, Guide, CRISPR-Cas Systems / genetics*
RNA, Guide, CRISPR-Cas Systems / metabolism
Viral Load / drug effects
Virus Latency / genetics
Virus Replication

Substances

Antineoplastic Agents
Bacterial Proteins
DNA, Viral
RNA, Guide, CRISPR-Cas Systems
CRISPR-Associated Protein 9
Cas9 endonuclease Streptococcus pyogenes
Endonucleases
Cisplatin
Fluorouracil