Harnessing the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated Cas9 system to disrupt the hepatitis B virus

The current therapies to treat hepatitis B virus (HBV) infection are limited. Recently, clustered regularly interspaced short palindromic repeat (CRISPR) systems, originally identified in bacteria and archaea, have been found to consist of an RNA-based adaptive immune system that degrades compliment...

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Bibliographic Details
Published in:Gene therapy 2015-05, Vol.22 (5), p.404-412
Main Authors: Zhen, S, Hua, L, Liu, Y-H, Gao, L-C, Fu, J, Wan, D-Y, Dong, L-H, Song, H-F, Gao, X
Format: Article
Language:English
Subjects:
Animals
Archaea
Care and treatment
Causes of
Cell culture
CRISPR
CRISPR-Cas Systems
Culture media
Deoxyribonucleic acid
DNA
Enzyme-linked immunosorbent assay
Enzymes
Female
Genetic Therapy
Hep G2 Cells
Hepatitis
Hepatitis B
Hepatitis B - therapy
Hepatitis B surface antigen
Hepatitis B Surface Antigens - genetics
Hepatitis B virus
Hepatitis B virus - genetics
Hepatocytes
Humans
Immune system
Immunohistochemistry
Lamivudine
Liver diseases
Mice
Mice, Inbred BALB C
Mutation
Pathogenic microorganisms
Plasmids
Ribonucleic acid
Risk factors
RNA
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Summary:The current therapies to treat hepatitis B virus (HBV) infection are limited. Recently, clustered regularly interspaced short palindromic repeat (CRISPR) systems, originally identified in bacteria and archaea, have been found to consist of an RNA-based adaptive immune system that degrades complimentary sequences of invading plasmids and viruses. Here, we studied the effects of the CRISPR/CRISPR-associated Cas9 system that was targeted to the surface antigen (HBsAg)-encoding region of HBV, both in a cell culture system and in vivo. The HBsAg levels in the media of the cells and in the sera of mice were analyzed by a quantitative enzyme-linked immunosorbent assay. The HBV DNA levels were assessed by quantitative PCR and HBsAg expression in mouse livers was assessed by an immunohistochemical assay. The amount of HBsAg secreted in the cell culture and mouse serum was reduced by CRISPR/Cas9 treatment. Immunohistochemistry analyses showed almost no HBsAg-positive cells in the liver tissue of CRISPR/Cas9-S1+X3-treated mice. The CRISPR/Cas9 system efficiently produced mutations in HBV DNA. Thus, CRISPR/Cas9 inhibits HBV replication and expression in vitro and in vivo and may constitute a new therapeutic strategy for HBV infection.
ISSN:0969-7128
1476-5462
DOI:10.1038/gt.2015.2