Suppression of hepatitis B virus DNA accumulation in chronically infected cells using a bacterial CRISPR/Cas RNA-guided DNA endonuclease

被引：191

作者：

Kennedy, Edward M. ^{[1
,2
]}

Bassit, Leda C. ^{[3
]}

Mueller, Henrik ^{[4
]}

Kornepati, Anand V. R. ^{[1
,2
]}

Bogerd, Hal P. ^{[1
,2
]}

Nie, Ting ^{[3
]}

Chatterjee, Payel ^{[3
]}

Javanbakht, Hassan ^{[4
]}

Schinazi, Raymond F. ^{[3
]}

Cullen, Bryan R. ^{[1
,2
]}

机构：

[1] Duke Univ, Med Ctr, Dept Mol Genet & Microbiol, Durham, NC USA

[2] Duke Univ, Med Ctr, Ctr Virol, Durham, NC USA

[3] Emory Univ, Sch Med, Dept Pediat, Ctr AIDS Res,Biochem Pharmacol Lab, Atlanta, GA 30322 USA

[4] F Hoffmann LaRoche Inc, Infect Dis, Basel, Switzerland

来源：

VIROLOGY | 2015年 / 476卷

关键词：

HBV; cccDNA; CRISPR/Cas; Antiviral; DNA editing; Lentiviral vector; HIGH-THROUGHPUT; SYSTEM; GENE; HBV; PERSISTENCE; LATENT; CCCDNA; ASSAY; STEP;

D O I：

10.1016/j.virol.2014.12.001

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Hepatitis B virus (HBV) remains a major human pathogen, with over 240 million individuals suffering from chronic HBV infections. These can persist for decades due to the lack of therapies that can effectively target the stable viral covalently closed circular (ccc) DNA molecules present in infected hepatocytes. Using lentiviral transduction of a bacterial Cas9 gene and single guide RNAs (sgRNAs) specific for HBV, we observed effective inhibition of HBV DNA production in in vitro models of both chronic and de novo HBV infection. Cas9/5gRNA combinations specific for HBV reduced total viral DNA levels by up to similar to 1000-fold and HBV cccDNA levels by up to similar to 10-fold and also mutationally inactivated the majority of the residual viral DNA. Together, these data provide proof of principle for the hypothesis that CRISPR/Cas systems have the potential to serve as effective tools for the depletion of the cccDNA pool in chronically HBV infected individuals. (C) 2014 Elsevier Inc. All rights reserved.

引用

页码：196 / 205

页数：10

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