CRISPR/Cas9 cleavage of viral DNA efficiently suppresses hepatitis B virus

Vyas Ramanan; Amir Shlomai; David B T Cox; Robert E Schwartz; Eleftherios Michailidis; Ankit Bhatta; David A Scott; Feng Zhang; Charles M Rice; Sangeeta N Bhatia

doi:10.1038/srep10833

CRISPR/Cas9 cleavage of viral DNA efficiently suppresses hepatitis B virus

Sci Rep. 2015 Jun 2:5:10833. doi: 10.1038/srep10833.

Authors

Affiliations

¹ Department of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
² Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, NY 10065, USA.
³ 1] Department of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA [2] Broad Institute, Cambridge, MA 02139, USA [3] Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.
⁴ 1] Department of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA [2] Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA [3] Division of Gastroenterology and Hepatology, Weill Cornell Medical College, New York, NY 10065, USA.
⁵ 1] Broad Institute, Cambridge, MA 02139, USA [2] McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139.
⁶ 1] Department of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA [2] Broad Institute, Cambridge, MA 02139, USA [3] Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139 [4] McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139.
⁷ 1] Department of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA [2] Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA [3] Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA [4] Broad Institute, Cambridge, MA 02139, USA [5] Howard Hughes Medical Institute, Cambridge, MA 02139, USA [6] Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

Abstract

Chronic hepatitis B virus (HBV) infection is prevalent, deadly, and seldom cured due to the persistence of viral episomal DNA (cccDNA) in infected cells. Newly developed genome engineering tools may offer the ability to directly cleave viral DNA, thereby promoting viral clearance. Here, we show that the CRISPR/Cas9 system can specifically target and cleave conserved regions in the HBV genome, resulting in robust suppression of viral gene expression and replication. Upon sustained expression of Cas9 and appropriately chosen guide RNAs, we demonstrate cleavage of cccDNA by Cas9 and a dramatic reduction in both cccDNA and other parameters of viral gene expression and replication. Thus, we show that directly targeting viral episomal DNA is a novel therapeutic approach to control the virus and possibly cure patients.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
CRISPR-Associated Proteins / genetics
CRISPR-Associated Proteins / metabolism
CRISPR-Cas Systems*
Cell Line, Tumor
Cells, Cultured
Clustered Regularly Interspaced Short Palindromic Repeats / genetics
DNA, Circular
DNA, Viral / genetics*
DNA, Viral / metabolism*
Disease Models, Animal
Gene Expression Regulation, Viral
Gene Targeting
Hepatitis B / virology
Hepatitis B virus / genetics*
Humans
Mice
RNA, Guide, CRISPR-Cas Systems
Virus Replication / genetics*

Substances

CRISPR-Associated Proteins
DNA, Circular
DNA, Viral
RNA, Guide, CRISPR-Cas Systems

Abstract

Publication types

MeSH terms

Substances

Grants and funding